Water-soluble amide derivatives of polyene macrolides and preparation and uses thereof

ABSTRACT

The present invention provides two new classes of polyene macrolide amide derivatives useful for treating or preventing fungal infections. The new polyene macrolide amide derivatives exhibit antifungal activity and are more water-soluble than conventional polyene antibiotics, such as amphotericin B and amphotericin B methyl ester.

1. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U.S.C. § 119(e) to U.S.provisional application Serial No. 60/207,659, filed May 31, 2000, thedisclosure of which is incorporated herein by reference in its entirety.

2. BACKGROUND OF THE INVENTION

[0002] 2.1 Field of the Invention

[0003] The present invention relates generally to derivatives of polyenemacrolides. In particular, the present invention relates towater-soluble amide derivatives of polyene macrolides useful fortreating or preventing topical and/or systemic fungal infections inplants, humans and animals.

[0004] 2.2 Description of Related Art

[0005] Many polyene macrolides are known that have antifungal propertiesuseful for treating topical and/or systemic fungal infections. Examplesof these polyene macrolides include amphotericin B, aureofacin,candicidin, candidin, levorin, mycoheptin, nystatin, partricin A,partricin B, perimycin, pimaricin, polyfungin, rimocidin andtrichomycin. However, due to their amphoteric character, these compoundsgenerally have limited solubility in aqueous solutions, which in turnlimits their usefulness in the treatment of systemic fungal infections.In prior attempts to modify these compounds, some of the derivativesexhibited undesirable toxic properties when used systemically. Forexample, while amphotericin B methyl ester (AME) exhibited lower acute,nephro- and hepato-toxicity than amphotericin B in rats and dogs, in theonly clinical trial conducted with AME patents with systemic fungalinfections, many patients developed progressive neurological dysfunctionassociated with white matter degeneration, see Ellis et al., 1988, ToxPath. 16(1):1; Parmegiani et al., 1987, Antimicrob. Agents Chemo.31(11):1756-60; Hoeprich et al., 1985, Diag. Microbiol. Infect. Dis.3:47-58; Massa et al., 1985, Fund. App. Tox. 5:737-753; Keim Jr., etal., 1976, Antimicrob. Agents Chemo. 10(4):687-690; and Keim Jr., etal., 1973, Science 179:584-586. The incidence and severity of thesecomplications increased with the amount of AME administered (Id.). Infact, the toxicity of AME was so severe that the clinical trial wascanceled and the product was never brought to market.

[0006] Many derivatives of polyene macrolides have been developed, inpart to address these limitations. One class of derivatives includecertain polyene macrolides substituted at the amino group of the aminosugar residue. For example, U.S. Pat. No. 4,093,796 to Falkowski et al.teaches polyene macrolides substituted at the sugar amino group with asaccharide. U.S. Pat. No. 4,195,172 to Falkowski et al. teachesN-methylglucamine salts of N-glycosyl derivatives of polyene macrolidesin which the amino group of the polyene macrolide is substituted with analdose or ketose mono- or oligosaccharide. U.S. Pat. No. 4,294,958 toFalkowski et al. teaches trimethylammonium salts of polyene macrolides,including the methyl esters. U.S. Pat. No. 4,365,058 to Falkowski et al.teaches esters of polyene macrolides that are substituted at the sugaramino group with non-sugar substituents. U.S. Pat. No. 5,314,999 toSeman et al. teaches polyene macrolides substituted at the N positionwith a 1-amino-1-deoxyketose group, which itself may be furthersubstituted. U.S. Pat. No. 5,942,495 to Borowski et al. teachesN-alkyl-N-glycosyl derivatives of polyene macrolides that are reportedto have antifungal activity, form water-soluble salts with acids, andhave lower toxicity than other N-alkyl polyene macrolide derivatives.

[0007] Other derivatives reported in the literature include amides ofcertain polyene macrolides derivatives. For example, U.S. Pat. No.4,783,527 to Falkowski et al. teaches alkyl, isoalkyl and heterocyclicamide derivatives of polyene macrolides. Jarzbeslo et al., 1982, J.Antibiot. 35(2):220-229 teach aliphatic amides of amphotericin B.Czerwinski et al., 1990, J. Antibiot. 43(6):680-683 teach amphotericin B2-morpholinoethylamide. Grzybowska & Borowski, 1990, J. Antibiot.43(7):907-908 teach hydrazides of amphotericin B, candidin, aureofacinand nystatin. Graybill et al., 1998, Antimicrobial Agents and Chemother.42(1)147-150 and Yamashita et al., 1995, J. Am. Chem. Soc.117(23):6249-6253 teach oligo(ethlene glycol) amides of amphotericin B.Cheron et al., 1988, Biochem. Pharmacol. 37(5):827-836 teach certainalkyl amides of amphotericin B that are further amidated at the polyeneamino sugar residue. Lastly, Bruzzese et al., 1996, Eur. J. Med. Chem.31:965-972, U.S. Pat. No. 5,296,597 to Bruzzese et al. and U.S. Pat. No.5,298,495 to Bruzzese et al. teach certain amide derivatives ofpartricins A and B.

[0008] None of the foregoing derivatives provide an optimum combinationof water solubility, low toxicity, and potency as an antifungal agent.Since AmB is still the drug of choice for many indications, there is aneed for polyene macrolide derivatives that exhibit antifungal activityand that have improved water solubility and/or toxicity properties.

3. SUMMARY OF THE INVENTION

[0009] In one aspect, the present invention provides new polyenemacrolide amide derivatives that have antifungal activity and that haveincreased water solubility as compared with amphotericin B (AmB) andamphotericin B methyl ester (AME). The polyene macrolide amidederivatives generally comprise a “core” polyene macrolide backbonederived from any of a variety of parent polyene macrolides having twofeatures: an exocyclic carboxyl group and an amino sugar residue. Theexocyclic carboxyl group of the parent polyene macrolide is amidatedwith substituents that increase the water-solubility of the resultantpolyene as compared with AmB and AME. In one class of polyene macrolideamide derivatives of the invention, the nitrogen atom of the primaryamino group of the amino sugar residue (“amino nitrogen”) is substitutedwith a carbohydrate residue, which may be a mono-, di-, oligo- orpolysaccharide. In all of the compounds of the invention, the aminonitrogen may be optionally alkylated. In embodiments in which the aminonitrogen is dialkylated, the alkyl groups may be the same or different.

[0010] While not intending to be bound by any particular theory ofoperation, the increased water-solubility of the amide derivatives ofthe invention is believed to be due to the presence of one or more ofthe same or different water solubility-increasing substituents attachedto and/or including the nitrogen atom of the amide group (“amidenitrogen”). The water-solubility increasing substituents are generallypolar in character, typically by virtue of including one or more of thesame or different substituted or unsubstituted heteroatoms (e.g., S, O,N, NH, etc.).

[0011] In one embodiment, the water-solubility increasing substituentsare hydrocarbons such as, by way of example and not limitation, linearand branched alkyls, cycloalkyls, aryls and arylalkyls that aresubstituted with one or more of the same or different polarsubstituents. Typical polar substituents include, but are not limitedto, —OH, —SH, ═O (oxo), ═S (thioxo), —NH₂, ═NH (imino), —C(═NH)—NH₂(amidino), —NH—C(═NH)—NH₂ (guanidino), —C(O)H, —C(O)OH, —C(O)O⁻M⁺,—C(O)NH₂, —N₃, —CN, —X, —CX₃, etc., where each X is independently ahalogen, preferably F, Cl or Br and M⁺ represents a monovalent counterion such as Na⁺, K⁺, etc. The polar-substituted alkyls, cycloalkyls,aryls and arylalkyls may also be optionally substituted with one or moreof the same or different non-polar substituents, e.g., alkyls,cycloalkyls, aryls and arylalkyls, etc.

[0012] In another embodiment, the water-solubility increasingsubstituents are hydrocarbons in which one or more of the carbon atomsare replaced with the same or different heteroatoms to form, by way ofexample and not limitation, linear and branched heteroalkyls,cycloheteroalkyls, heteroaryls and heteroarylalkyls. One or more of thecarbon atoms and/or heteroatoms (e.g., N) of these heteroalkyl,cycloheteroalkyl, heteroaryl and heteroarylalkyl groups may be furthersubstituted with one or more of the same or different polar or non-polarsubstituents, as described above.

[0013] In still another embodiment, the water solubility-increasingsubstituents, taken together with the amide nitrogen to which they arebonded, form a saturated or unsaturated nitrogen-containing ring. Thering may optionally include one or more of the same or differentadditional ring heteroatoms, and/or may be optionally substituted at oneor more ring carbon or heteroatoms with the same or different polar ornon-polar substituents, as previously described.

[0014] In one embodiment of the invention, the watersolubility-increasing substituents are selected from the groupconsisting of polyhydroxylated alkyls, mono-, di-, oligo- andpolysaccharides, polyalkylene glycols (e.g., polyethylene glycol,polypropylene glycol, etc.) and polyalkylene oxides.

[0015] In embodiments of the invention in which the amide nitrogenincludes only a single water-solubility increasing substituent, theother amide nitrogen substituent may be a hydrogen or a non-polarsubstituent, as described above.

[0016] In embodiments of the invention which include a carbohydrateresidue at the amino nitrogen, the carbohydrate residue is generallyadded via an Amadori rearrangement with a reducing carbohydrate.

[0017] In one illustrative embodiment, the present invention providespolyene macrolide amide derivatives according to structural formula (I):

[0018] including the pharmaceutically acceptable salts thereof, wherein:

[0019] N—R¹—C(O) is a polyene macrolide backbone;

[0020] CH₂—R² is a carbohydrate residue, where the illustrated CH₂ isderived from the anomeric carbon of a terminal carbohydrate saccharideand R² represents the remainder of the carbohydrate;

[0021] either: (i) R⁶ and R⁷ are each, independently of one another,selected from the group consisting of hydrogen, non-polar substituentand water-solubility increasing substituent, with the proviso that atleast one of R⁶ or R⁷ is a water-solubility increasing substituent; or(ii) R⁶ and R⁷, taken together with the amide nitrogen to which they arebonded, form a saturated or unsaturated ring which optionally includesone or more of the same or different additional ring heteroatoms andwhich is optionally further substituted with one or more of the same ordifferent polar or non-polar substituent or combinations thereof; and

[0022] R¹⁴ is hydrogen or alkyl.

[0023] In another illustrative embodiment, the present inventionprovides polyene macrolide amide derivatives according to structuralformula (II):

[0024] including the pharmaceutically acceptable salts thereof, wherein:

[0025] N—R¹—C(O) and R¹⁴ are as previously defined for structuralformula (I);

[0026] R³ is hydrogen, a non-polar substituent or a water-solubilityincreasing substituent;

[0027] R⁴ is hydrogen or alkyl; and

[0028] R⁵ is a water-solubility increasing substituent selected from thegroup consisting of polyhydroxylated alkyl, monosaccharide, disaccharideand oligosaccharide.

[0029] In certain embodiments of the polyene macrolide amide derivativesaccording to structural formula (I), R⁶ is hydrogen and/or R¹⁴ ishydrogen. In certain embodiments of the polyene macrolide amidederivatives according to structural formula (II), R³ is hydrogen and/orone or both of R⁴ and R¹⁴ are hydrogen.

[0030] In another aspect, the present invention provides methods ofmaking the polyene macrolide amide derivatives of the invention. Toobtain an amide derivative according to structural formula (II) in whichR⁴ and R¹⁴ are each hydrogen, a parent polyene macrolide is reacted withan appropriate amine according to known methods or the methods describedbelow. Derivatives according to structural formula (II) in which R⁴and/or R¹⁴ is alkyl may be obtained from the above product usingstandard alkylation methods. Alternatively, the parent polyene macrolidemay be first alkylated according to standard methods and the alkylatedintermediate amidated according to known methods or the methodsdescribed below.

[0031] To obtain a polyene macrolide amide derivative according tostructural formula (I) in which R¹⁴ is hydrogen, a parent polyenemacrolide is first reacted with an appropriate reducing carbohydrateunder Amadori rearrangement conditions to yield an Amadori product thatis substituted at the amino nitrogen with a carbohydrate residue. ThisAmadori product is then reacted with an appropriate amine according toknown methods or the methods described below to yield a polyene amidederivative according to structural formula (I). Derivatives according tostructural formula (I) in which R¹⁴ is alkyl may be obtained from theabove product using standard alkylation methods. Alternatively, theparent polyene macrolide may be first monoalkylated prior to the Amadorirearrangement reaction, or the Amadori product may be alkylated prior tothe amidation reaction.

[0032] Alternatively, the amide derivatives according to structuralformula (I) may be prepared by reacting an amide derivative according tostructural formula (II) in which at least one of R⁴ or R¹⁴ is hydrogenwith an appropriate reducing carbohydrate under Amadori rearrangementconditions. If this alternative route is used, any substituents on theamide moiety that are capable of reacting with the reducing carbohydrateshould be protected prior to reaction with the reducing carbohydrate. IfR⁴ and R¹⁴ of the starting material were each hydrogen, derivativesaccording to structural formula (I) in which R¹⁴ is alkyl may beobtained from the above product using standard alkylation methods.

[0033] In a specific embodiment of the invention, the amidation step ofthe synthesis is carried out using a uronium salt or phosphonium saltcoupling reagent. The reaction is typically carried out in the presenceof an organic amine-containing base.

[0034] In another embodiment, the polyene macrolide amide derivatives offormula (I) are synthesized in a one-pot reaction. According to thisembodiment, a parent polyene macrolide is first reacted with a reducingcarbohydrate under Amadori rearrangement conditions. Without anyisolation or purification, the Amadori product is then amidatedutilizing a uronium salt or phosphonium salt coupling reagent, asdescribed above. If an alkylated derivative is desired, the parentpolyene macrolide should be alkylated first and then used as a startingreagent in the one-pot reaction.

[0035] In all of the described synthetic routes, where applicable, theintermediates and/or reaction products may be isolated using standardtechniques, such as precipitation and/or chromatography.

[0036] In another aspect, the present invention provides pharmaceuticalcompositions including the new polyene macrolide amide derivatives. Thepharmaceutical compositions generally comprise one or more polyenemacrolide amide derivatives of the invention (which may be in the formof the pharmaceutically acceptable salts thereof) and a pharmaceuticallyacceptable carrier, excipient or diluent. The choice of carrier,excipient or diluent will depend upon, among other factors, the desiredmode of administration.

[0037] In still another aspect the present invention provides methods ofinhibiting the growth of fungi, such as C. albicans and other Candidaspecies (e.g., C. glabrata), Crytococcus neoformans, Blastomycesdermatitidis, Histoplasma capsulatum, Torulopsis glabrata, Coccidioidesimmitus, Paracoccidioides braziliensis, Aspergillus species and theagents of mucormycosis. The method generally involves contacting afungus with an amount of a polyene macrolide amine derivative of theinvention, or a pharmaceutically-acceptable salt thereof, effective toinhibit the growth of the fungus. The method may be practiced to achievea fungistatic effect, where the growth of the fungus is inhibited, or toachieve a fungicidal effect, where the fungus is killed.

[0038] In a final aspect, the present invention provides methods fortreating and/or preventing fungal infections in humans, animals and/orplants. The methods generally involve administering to a human, animalor plant one or more of the polyene macrolide amide derivatives orpharmaceutical compositions of the invention in an amount effective totreat or prevent a fungal infection in the human, animal or plant. Thepolyene macrolide amide derivatives or pharmaceutical compositions maybe administered systemically or applied topically, depending on thenature of the fungal infection.

4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0039] 4.1 Definitions

[0040] As used herein, the following terms are intended to have thefollowing meanings.

[0041] “Alkyl” refers to a saturated or unsaturated, branched,straight-chain or cyclic monovalent hydrocarbon group derived by theremoval of one hydrogen atom from a single carbon atom of a parentalkane, alkene or alkyne. Typical alkyl groups include, but are notlimited to, methyl; ethyls such as ethanyl, ethenyl, ethynyl; propylssuch as propan-1-yl, propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl,prop-1-en-2-yl, prop-2-en-1-yl (allyl), cycloprop-1-en-1-yl;cycloprop-2-en-1-yl, prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls suchas butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl,cyclobutan-1-yl, but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl,but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

[0042] The term “alkyl” is specifically intended to include groupshaving any degree or level of saturation, i.e., groups havingexclusively carbon-carbon single bonds, groups having one or morecarbon-carbon double bonds, groups having one or more carbon-carbontriple bonds and groups having mixtures of single, double and triplecarbon-carbon bonds. Where a specific level of saturation is intended,the expressions “alkanyl,” “alkenyl,” and “alkynyl” are used. Theexpression “lower alkyl” refers to alkyl groups comprising from 1 to 8carbon atoms.

[0043] The number of carbon atoms comprising a particular alkyl may varywidely, and is limited only by the properties of the resultant molecule.For example, if an alkyl is included as a non-polar substituent group ona water-solubility increasing substituent, it should have a number ofcarbon atoms that do not cause the water-solubility increasingsubstituent to become net hydrophobic in character. In this instance,the alkyl will generally comprise from 1 to 20 carbon atoms, typicallyfrom 1 to 10 carbon atoms, usually from 1 to 8 carbon atoms and mostfrequently from 1 to 4 carbon atoms. For amide derivatives on theinvention that are alkylated at the amino nitrogen, the alkyl willgenerally comprise from 1 to 20 carbon atoms, typically from 1 to 10carbon atoms, usually from 1 to 8 carbon atoms and most frequently from1 to 4 carbon atoms, although it may comprise greater numbers of carbonatoms provided that the resultant molecule is active as describedherein.

[0044] “Alkanyl” refers to a saturated branched, straight-chain orcyclic alkyl group derived by the removal of one hydrogen atom from asingle carbon atom of a parent alkane. Typical alkanyl groups include,but are not limited to, methanyl; ethanyl; propanyls such aspropan-1-yl, propan-2-yl (isopropyl), cyclopropan-1-yl, etc.; butyanylssuch as butan-1-yl, butan-2-yl (sec-butyl), 2-methyl-propan-1-yl(isobutyl), 2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, etc.; andthe like.

[0045] “Alkenyl” refers to an unsaturated branched, straight-chain orcyclic alkyl group having at least one carbon-carbon double bond derivedby the removal of one hydrogen atom from a single carbon atom of aparent alkene. The group may be in either the cis or trans conformationabout the double bond(s). Typical alkenyl groups include, but are notlimited to, ethenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl,prop-2-en-1-yl (allyl), prop-2-en-2-yl, cycloprop-1-en-1-yl;cycloprop-2-en-1-yl; butenyls such as but-1-en-1-yl, but-1-en-2-yl,2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl,buta-1,3-dien-1-yl, buta-1,3-dien-2-yl, cyclobut-1-en-1-yl,cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, etc.; and the like.

[0046] “Alkynyl” refers to an unsaturated branched, straight-chain orcyclic alkyl group having at least one carbon-carbon triple bond derivedby the removal of one hydrogen atom from a single carbon atom of aparent alkyne. Typical alkynyl groups include, but are not limited to,ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1-yl, etc.;butynyls such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; andthe like.

[0047] “Cycloalkyl” refers to a saturated or unsaturated cyclic alkylgroup. Where a specific level of saturation is intended, thenomenclature “cycloalkanyl” or “cycloalkenyl” is used. Typicalcycloalkyl groups include, but are not limited to, groups derived fromcyclopropane, cyclobutane, cyclopentane, cyclohexane, and the like. In apreferred embodiment, the cycloalkyl group is (C₃-C₆) cycloalkyl, morepreferably (C₃-C₆) cycloalkanyl.

[0048] “Alkyldivl” refers to a saturated or unsaturated, branched,straight-chain or cyclic divalent hydrocarbon group derived by theremoval of one hydrogen atom from each of two different carbon atoms ofa parent alkane, alkene or alkyne, or by the removal of two hydrogenatoms from a single carbon atom of a parent alkane, alkene or alkyne.The two monovalent radical centers or each valency of the divalentradical center can form bonds with the same or different atoms. Typicalalkyldiyl groups include, but are not limited to methandiyl; ethyldiylssuch as ethan-1,1-diyl, ethan-1,2-diyl, ethen-1,1-diyl, ethen-1,2-diyl;propyldiyls such as propan-1,1-diyl, propan-1,2-diyl, propan-2,2-diyl,propan-1,3-diyl, cyclopropan-1,1-diyl, cyclopropan-1,2-diyl,prop-1-en-1,1-diyl, prop-1-en-1,2-diyl, prop-2-en-1,2-diyl,prop-1-en-1,3-diyl, cycloprop-1-en-1,2-diyl, cycloprop-2-en-1,2-diyl,cycloprop-2-en-1,1-diyl, prop-1-yn-1,3-diyl, etc.; butyldiyls such as,butan-1,1-diyl, butan-1,2-diyl, butan-1,3-diyl, butan-1,4-diyl,butan-2,2-diyl, 2-methyl-propan-1,1-diyl, 2-methyl-propan-1,2-diyl,cyclobutan-1,1-diyl; cyclobutan-1,2-diyl, cyclobutan-1,3-diyl,but-1-en-1,1-diyl, but-1-en-1,2-diyl, but-1-en-1,3-diyl,but-1-en-1,4-diyl, 2-methyl-prop-1-en-1,1-diyl,2-methanylidene-propan-1,1-diyl, buta-1,3-dien-1,1-diyl,buta-1,3-dien-1,2-diyl, buta-1,3-dien-1,3-diyl, buta-1,3-dien-1,4-diyl,cyclobut-1-en-1,2-diyl, cyclobut-1-en-1,3-diyl, cyclobut-2-en-1,2-diyl,cyclobuta-1,3-dien-1,2-diyl, cyclobuta-1,3-dien-1,3-diyl,but-1-yn-1,3-diyl, but-1-yn-1,4-diyl, buta-1,3-diyn-1,4-diyl, etc.; andthe like. Where specific levels of saturation are intended, thenomenclature alkanyldiyl, alkenyldiyl and/or alkynyldiyl is used. Inpreferred embodiments, the alkyldiyl group is (C₁-C₆) alkyldiyl. Alsopreferred are saturated acyclic alkanyldiyl groups in which the radicalcenters are at the terminal carbons, e.g., methandiyl (methano);ethan-1,2-diyl (ethano); propan-1,3-diyl (propano); butan-1,4-diyl(butano); and the like (also referred to as alkylenos, defined infra).

[0049] “Alkyleno” refers to a straight-chain alkyldiyl group having twoterminal monovalent radical centers derived by the removal of onehydrogen atom from each of the two terminal carbon atoms ofstraight-chain parent alkane, alkene or alkyne. Typical alkyleno groupsinclude, but are not limited to, methano; ethylenos such as ethano,etheno, ethyno; propylenos such as propano, prop[1]eno, propa[1,2]dieno,prop[1]yno, etc.; butylenos such as butano, but[1]eno, but[2]eno,buta[1,3]dieno, but[1]yno, but[2]yno, but[1,3]diyno, etc.; and the like.Where specific levels of saturation are intended, the nomenclaturealkano, alkeno and/or alkyno is used. In preferred embodiments, thealkyleno group is (C₁-C₆) or (C₁-C4) alkyleno. Also preferred arestraight-chain saturated alkano groups, e.g., methano, ethano, propano,butano, and the like.

[0050] “Heteroalkyl, Heteralkanyl, Heteroalkenyl, Heteroalkanyl,Heteroalkyldivl and Heteroalkyleno” refer to alkyl, alkanyl, alkenyl,alkynyl, alkyldiyl and alkyleno groups, respectively, in which one ormore of the carbon atoms (and any associated hydrogen atoms) are eachindependently replaced with the same or different heteroatoms orheteroatomic groups. Typical heteroatoms or heteroatomic groups whichcan be included in these groups include, but are not limited to, —O—,—S—, —Se—, —O—O—, —S—S—, —O—S—, —O—S—O—, —O—NR′—, —NR′—, —NR′—NR′—,═N—N═, —N═N—, —N═N—NR′—, —PH—, —P(O)₂—, —O—P(O)₂—, —SH₂, —S(O)₂—, —SnH₂—and the like, and combinations thereof, including, for example,—NR′—S(O)₂—, where each R′ is independently selected from the groupconsisting of hydrogen, alkyl, alkanyl, alkenyl, alkynyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl, as defined herein.

[0051] “Cycloheteroalkyl” refers to a saturated or unsaturated cyclicalkyl group in which one or more carbon atoms (and any associatedhydrogen atoms) are independently replaced with the same or differentheteroatom or heteroatomic group. Typical heteroatoms to replace thecarbon atom(s) include, but are not limited to, N, P, O, S, Si, etc.(including any associated hydrogen atoms, e.g., NH). Where a specificlevel of saturation is intended, the nomenclature “cycloheteroalkanyl”or “cycloheteroalkenyl” is used. Typical cycloheteroalkyl groupsinclude, but are not limited to, groups derived from epoxides,imidazolidine, morpholine, piperazine, piperidine, pyrazolidine,pyrrolidine, quinuclidine, and the like. In preferred embodiments, thecycloheteroalkyl is a 3-6 membered cycloheteroalkyl. Particularlypreferred cycloheteralkyls are morpholino, pyrrolidino, pipyridino,tetrahydrothiopheno, tetrahydrofuranyl and tetrahydropyranyl.

[0052] “Parent Aromatic Ring System” refers to an unsaturated cyclic orpolycyclic ring system having a conjugated π electron system.Specifically included within the definition of “parent aromatic ringsystem” are fused ring systems in which one or more of the rings arearomatic and one or more of the rings are saturated or unsaturated, suchas, for example, indane, indene, phenalene, etc. Typical parent aromaticring systems include, but are not limited to, aceanthrylene,acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene,hexalene, as-indacene, s-indacene, indane, indene, naphthalene,octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene,pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene,and the like.

[0053] “Aryl” refers to a monovalent aromatic hydrocarbon group derivedby the removal of one hydrogen atom from a single carbon atom of aparent aromatic ring system. Typical aryl groups include, but are notlimited to, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene, and the like. In preferredembodiments, the aryl group is (C₅-C₁₄) aryl, with (C₅-C₁₀) being evenmore preferred. Particularly preferred aryls are cyclopentadienyl,phenyl and naphthyl.

[0054] “Arylalkyl” refers to an acyclic alkyl group in which one of thehydrogen atoms bonded to a carbon atom, typically a terminal or sp³carbon atom, is replaced with an aryl group. Typical arylalkyl groupsinclude, but are not limited to, benzyl, 2-phenylethan-1-yl,2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethan-1-yl,2-naphthylethen-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl and thelike. Where specific alkyl moieties are intended, the nomenclaturearylalkanyl, arylakenyl and/or arylalkynyl is used. In preferredembodiments, the arylalkyl group is (C₆-C₂₀) arylalkyl, e.g., thealkanyl, alkenyl or alkynyl moiety of the arylalkyl group is (C₁-C₆) andthe aryl moiety is (C₅-C₁₄). In particularly preferred embodiments thearylalkyl group is (C₆-C₁₃), e.g., the alkanyl, alkenyl or alkynylmoiety of the arylalkyl group is (C₁-C₃) and the aryl moiety is(C₅-C₁₀).

[0055] “Parent Heteroaromatic Ring System” refers to a parent aromaticring system in which one or more carbon atoms (and any associatedhydrogen atoms) are each independently replaced with the same ordifferent heteroatoms. Typical heteratoms to replace the carbon atomsinclude, but are not limited to, N, P, O, S, Si, etc. (including anyassociated hydrogen atoms, e.g., NH). Specifically included within thedefinition of “parent heteroaromatic ring systems” are fused ringsystems in which one or more of the rings are aromatic and one or moreof the rings are saturated or unsaturated, such as, for example,arsindole, chromane, chromene, indole, indoline, xanthene, etc. Typicalparent heteroaromatic ring systems include, but are not limited to,arsindole, carbazole, β-carboline, chromane, chromene, cinnoline, furan,imidazole, indazole, indole, indoline, indolizine, isobenzofuran,isochromene, isoindole, isoindoline, isoquinoline, isothiazole,isoxazole, naphthyridine, oxadiazole, oxazole, perimidine,phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine,pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and thelike.

[0056] “Heteroaryl” refers to a monovalent heteroaromatic group derivedby the removal of one hydrogen atom from a single atom of a parentheteroaromatic ring system. Typical heteroaryl groups include, but arenot limited to, groups derived from acridine, arsindole, carbazole,β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole,indole, indoline, indolizine, isobenzofuran, isochromene, isoindole,isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine,oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline,phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline,quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,thiophene, triazole, xanthene, and the like. In preferred embodiments,the heteroaryl group is a 5-14 membered heteroaryl, with 5-10 memberedheteroaryl being particularly preferred. The most preferred heteroarylgroups are those derived from thiophene, pyrrole, benzothiophene,benzofuran, indole, pyridine, quinoline, imidazole, oxazole andpyrazine.

[0057] “Heteroarylalkyl” refers to an acyclic alkyl group in which oneof the hydrogen atoms bonded to a carbon atom, typically a terminal orsp³ carbon atom, is replaced with a heteroaryl group. Where specificalkyl moieties are intended, the nomenclature heteroarylalkanyl,heteroarylakenyl and/or heterorylalkynyl is used. In preferredembodiments, the heteroarylalkyl group is a 6-20 memberedheteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety of theheteroarylalkyl is 1-6 membered and the heteroaryl moiety is a5-14-membered heteroaryl. In particularly preferred embodiments, theheteroarylalkyl is a 6-13 membered heteroarylalkyl, e.g., the alkanyl,alkenyl or alkynyl moiety is 1-3 membered and the heteroaryl moiety is a5-10 membered heteroaryl.

[0058] 4.2 The Compounds

[0059] The present invention provides two new classes of polyenemacrolide amide derivatives (and/or pharmaceutically acceptable saltsthereof), pharmaceutical compositions comprising the new polyenemacrolide amide derivatives, methods of making the new polyene macrolideamide derivatives, and methods of using the new polyene macrolide amidederivatives and/or pharmaceutical compositions to inhibit the growth offungi and/or to treat and/or prevent fungal infections in both plantsand animals, including humans.

[0060] The polyene macrolide amide derivatives of the invention providesignificant advantages over traditional polyene macrolide antifungals.For example, the polyene macrolide amide derivatives of the presentinvention are more water soluble, and many exhibit lower acute toxicity,than traditional polyene macrolide antifungals such as amphotericin B(AmB) and amphotericin B methyl ester (AME). Owing in part to theirwater-solubility, the polyene macrolide amide derivatives of theinvention do not require extensive formulation, making them extremelyeasy for use. Most dissolve readily in water in their free base forms,and all are readily soluble in water when prepared as pharmaceuticallyacceptable salts, such as aspartate salts. Thus, the amide derivativesof the invention may be stored dry and dissolved in water or otheraqueous vehicles just prior to use. This is in stark contrast to AmB,which is typically sold as a lipid formulation (e.g., FUNGIZONE,Briston-Meyers Squibb Co.).

[0061] One class of polyene macrolide amide derivatives according to theinvention includes compounds according to structural formula (I):

[0062] including the pharmaceutically acceptable salts thereof, wherein:

[0063] N—R¹—C(O) is a polyene macrolide backbone;

[0064] CH₅—R² is a carbohydrate residue, where the illustrated CH₂ isderived from the anomeric carbon of a terminal carbohydrate saccharideand R² represents the remainder of the carbohydrate;

[0065] either: (i) R⁶ is selected from the group consisting of hydrogen,non-polar substituent and water-solubility increasing substituent and R⁷is a water-solubility increasing substituent; or (ii) R⁶ and R⁷, takentogether with the amide nitrogen to which they are bonded, form asaturated or unsaturated ring which optionally includes one or more ofthe same or different additional ring heteroatoms and which isoptionally further substituted with one or more of the same or differentpolar or non-polar substituent or combinations thereof; and

[0066] R¹⁴ is hydrogen or alkyl.

[0067] Another class of polyene macrolide amide derivatives according tothe invention includes compounds according to structural formula (II):

[0068] including the pharmaceutically acceptable salts thereof, wherein:

[0069] N—R¹—C(O) and R¹⁴ are as previously defined for structuralformula (I);

[0070] R³ is hydrogen, a non-polar substituent or a water-solubilityincreasing substituent;

[0071] R⁴ is hydrogen or alkyl; and

[0072] R⁵ is a water-solubility increasing substituent selected from thegroup consisting of polyhydroxylated alkyl, monosaccharide, disaccharideand oligosaccharide.

[0073] The compounds of the invention are amide derivatives of “parent”polyene macrolides of a particular type. Specifically, the parentpolyene macrolides are of a type that have an exocyclic carboxyl groupand an amino sugar residue, as exemplified by, for example, AmB andnystatin. In the compounds according to structural formulae (I) and(II), the exocyclic carboxyl group of the parent polyene macrolide isconverted to an amide group. The nitrogen atom of this exocyclic amidegroup (“amide nitrogen”) is substituted with one or more of the same ordifferent substituents that increase the water-solubility of theresultant compound as compared with AmB and AME, as will be described inmore detail, below.

[0074] In the compounds according to structural formula (I), the primaryamino group of the amino sugar residue of the parent polyene macrolideis substituted with a carbohydrate residue. This carbohydrate residue,which is described in more detail below, is attached to the nitrogenatom of this primary amino group (“amino nitrogen”) via the anomericcarbon of a terminal saccharide unit. The compounds of structuralformulae (I) and (II) may also be alkylated at this amino nitrogen. Inembodiments of the compounds of formula (II) which are dialkylated, thealkyl groups may be the same or different.

[0075] In the polyene macrolide amide derivatives of formulae (I) and(II), polyene backbone N—R¹—C(O) may be derived from any known or laterdiscovered parent polyene macrolide having the features discussed above.Preferably, the parent polyene macrolide from which backbone N—R¹—C(O)is derived will have antifungal activity. Non-limiting examples ofparent polyene macrolides having the required features from whichpolyene backbone N—R¹—C(O) may be derived include, but are not limitedto, amphotericin A (AmA), amphotericin B (AmB), aureofacin, candicidin,candidin, levorin, mycoheptin, nystatin (including A₁), partricin(including A and B), pentamycin, perimycin, pimaricin, polyfungin,rimocidin, and trichomycin. Preferred classes of polyene backbonesN—R¹—C(O) are those derived from AmB and nystatin. The structures ofnystatin and AmB are as illustrated below, including citationsreferencing methods for obtaining these parent polyene macrolides (foreach parent polyene macrolide, the required exocyclic carboxyl group andthe amino primary group of the required amino sugar residue areillustrated in bold and indicated with arrows): AmB

Merck Index #620; U.S. Pat. No. 2,508,611; Nystatin A₁

Merck Index #6658; U.S. Pat. Nos. 2,832,719 and 3,517,100

[0076] In the compounds according to structural formulae (I) and (II),the illustrated N—R¹—C(O) portion of the molecule is contributed by theparent polyene macrolide. Those of skill in the art will recognize thatwhen R⁴ and/or R¹⁴ are hydrogen, these hydrogens are also contributed bythe parent polyene macrolide. It will further be appreciated thatpolyene backbone N—R¹—C(O) includes the amino sugar moiety that isattached to the macrocyclic portion of parent polyene macrolide (e.g.,the 3-amino-3,6-dideoxymannose of AmB and nystatin). This amino sugar,which is an inherent part of the parent polyene macrolide, is to bedistinguished from the carbohydrate residue CH₂R² of formula (I), whichis not contributed by the parent polyene macrolide and constitutes oneof the inventive features of certain amide derivatives of the invention.

[0077] As a specific example to clarify the nomenclature and compoundsdescribed herein, the polyene backbone N—R¹—C(O) derived from AmB isillustrated below, wherein the bold dashed lines indicate the atomswhich are bonded to the CH₂—R², NR⁶R⁷ and R¹⁴ substituents in the amidederivatives of formula (I) and the NR³R⁵, R⁴ and R¹⁴ substituents in theamide derivatives of formula (II):

[0078] The structures of polyene backbones N—R¹—C(O) derived from otherparent polyene macrolides will be apparent to those of skill in the art.

[0079] To further illustrate the compounds of the invention, the amidederivatives of structural formulae (I) and (II) in which N—R¹—C(O) is apolyene macrolide backbone derived from AmB are provided below asstructures (Ia) and (IIa), respectively: (Ia)

(IIa)

[0080] In the amide derivatives of structural formula (I), thecarbohydrate residue CH₂—R² may be any number of saccharide units inlength, and typically ranges from 1 to about 100 saccharide units. Thus,the carbohydrate residue can be a monosaccharide, a disaccharide, anoligosaccharide comprising from three to tens of saccharide units or apolysaccharide comprising from tens to 30, 40, 50, 60, 100, severalhundred, several thousand or even more, saccharide units. In mostinstances, the carbohydrate residue will be a mono-, di- oroligosaccharide. However, as it has been discovered that substitutingthe amino sugar residue of AmB with large polymers does notdeleteriously affect the antifungal activity of the compound,carbohydrate residue CH₂—R² may be a large, water-insolublepolysaccharide and still retain antifungal activity. Polyene macrolideamide derivatives including large polysaccharides that have lowwater-solubility for carbohydrate residue CH₂—R² may be used topicallyor as antifungals in non-aqueous environments. Alternatively, thewater-solubility characteristics of the compound may be improved byselecting a substituent R³ that has a high water-solubility, such as asubstituent that is highly polar, as will be discussed in more detailbelow. As the amide derivatives of formula (I) comprise water-solubilityincreasing substituents at the amide nitrogen, a significant advantageof the compounds of the invention is the ability to selectively tunetheir water-solubility through the choice of amide nitrogensubstituents.

[0081] The carbohydrate residue CH₂—R² may be a homopolymer, in whichall saccharide units are the same, or it may be a heteropolymercomprising mixtures of different saccharide units. The carbohydrateresidue may be branched or linear, and, as will be discussed in moredetail below, the saccharide units may be, independently of one another,in a cyclic conformation, a linear conformation or in a mixture ofcyclic and linear conformations. Moreover, subject only to theconstraints of the Amadori rearrangement reaction used to synthesize thepolyene macrolide amide derivatives of formula (I), the saccharide unitsof carbohydrate residue CH₂—R² may be substituted with a variety ofdifferent substituents. These substituents may be used to impart thederivatives of the invention with desirable properties, such as, forexample, improved water-solubility, lower toxicity, etc., and will beapparent to those of skill in the art.

[0082] As will be discussed in more detail in connection with themethods of synthesizing the amide derivatives of formula (I), it will beappreciated that carbohydrate residue CH₂—R² is produced via an Amadorirearrangement of an appropriate reducing carbohydrate, typically areducing sugar. Therefore, the carbohydrate residue CH₂—R² of thederivatives of formula (I) has a structure that is different from thereducing carbohydrate used as a reactant in the Amadori rearrangementreaction that yields the polyene macrolide amide derivatives of formula(I). The principles of the Amadori rearrangement reaction and therequirements of reducing carbohydrates that can undergo an Amadorirearrangement are well-known and well understood. Briefly, therearrangement and the requirements of the reducing carbohydrates,exemplified with a monosaccharide, are illustrated below:

Amadori Rearrangement

[0083]

[0084] The requirements of reducing carbohydrates which can undergo therearrangement are defined by the various R groups. Generally, thecarbohydrate reactant is an aldose, and the hydroxy group at the2-position must be present and unblocked. In compounds 40, 41, 42, 43,44 and 45, a is an integer from 0 up to virtually any number, where a is0 only in open chain conformers; R¹⁰ is hydrogen, alkyl, alkylidene,cycloalkyl, arylalkyl, aryl, glycosyl or polymer, but not acyl or astrongly electron-withdrawing radical; R¹¹ is hydrogen, alkyl,alkylidene, arylalkyl, but not aryl when R¹⁰ is aryl and, in addition,the combination of (R¹⁰+R¹¹) in the reacting amine should not stericallyhinder the nitrogen atom; and R¹³ is hydrogen, —CH₂OH, —CH₃, —COOH,—CONHR, —COO⁻M⁺, and the like, where R is, for example, hydrogen oralkyl and M⁺ represents a metal ion, such as Na⁺, K⁺, etc. Any reducingcarbohydrate having these attributes can undergo an Amadorirearrangement to yield the polyene macrolide amide derivatives offormula (I). For additional guidance regarding the requirements of theAmadori rearrangement, see Hodge & Fisher, “Amadori RearrangementProducts,” In: Methods in Carbohydrate Chemistry, Volume II, Reactionsof Carbohydrates, Whistler & Wolfram, Eds., pp. 99-107, Academic Press,Inc., New York (1963). Skilled artisans will be able to select anappropriate reducing carbohydrate reactant to obtain an amide derivativeaccording to formula (I) that has the desired carbohydrate residueCH₂—R². TABLE 1 presents an exemplary list of reducing carbohydratesthat are capable of undergoing an Amadori rearrangement that may be usedto produce polyene macrolide amide derivatives of structural formula(I). Other carbohydrates having appropriate properties will be apparentto those of skill in the art.

[0085] Those of skill in the art will appreciate that theabove-illustrated Amadori rearrangement illustrates only thecarbohydrate. For the reactions and polyene macrolide amide derivativesdescribed herein, in Compounds 40, 41, 42, 43, 44 and 45, R¹⁰corresponds to R¹—C(O) and R¹⁰ corresponds to R¹⁴ of structural formula(I). TABLE 1 Exemplary Reducing Carbohydrates for Amadori Reactionallose altrose arabinose cellobiose fucose galactose glucose3-O-methyl-glucose 4-fluoro-4-deoxy-glucose gulose idose lactose lyxosemaltopentose maltose mannoseN-(2,3,5-trihydroxy-6-hydroxymethyl-tetrahydro-pyran-4-yl)-acetamideribose talose xylose

[0086] The absolute stereochemistry of the carbohydrate residue CH₂—R²in the amide derivatives of structural formula (I) is not critical tosuccess. Thus, the reducing carbohydrates used as reactants, e.g., thereducing carbohydrates of TABLE 1, may be D-isomers, L-isomers ormixtures of D- and L-isomers, depending upon the desired stereochemistryof the resultant product. If optically pure compounds are desired,reducing carbohydrates that are pure optical isomers should be selectedas reactants for the Amadori rearrangement reaction.

[0087] The reducing carbohydrates, e.g., the reducing carbohydrates ofTABLE 1, may also be either α- or β-conformers, or mixtures of α- andβ-conformers. Owing to the mechanism of action of the Amadorirearrangement reaction, corresponding α- and β-conformer reducingcarbohydrate reactants will yield the same Amadori product.

[0088] As illustrated in the above rearrangement, it will be appreciatedthat the resultant carbohydrate residue produced by an Amadorirearrangement reaction may be in either a cyclic or linear conformation,or may be a mixture of cyclic and linear conformers. In the specificpolyene macrolide amide derivatives of formula (I) illustrated herein,the various carbohydrate residues added via the Amadori rearrangementare shown in their cyclic conformations. However, it will be appreciatedthat these illustrations are not intended in any way to limit thecarbohydrate residue of the illustrated derivatives, or of any polyenemacrolide amide derivatives according to structural formula (I), to thecyclic forms. When the carbohydrate residue is a monosaccharide, it maybe linear, cyclic or a mixture of linear and cyclic conformers. When thecarbohydrate residue is a di-, oligo- or polysaccharide, eachmonosaccharide unit may be cyclic or linear, or a mixture of cyclic andlinear conformers. Thus, the polyene macrolide amide derivativesaccording to structural formula (I) may be in the form of pure compoundsor in the form of mixtures of two or more different conformers. The onlyrequirement is that the polyene macrolide amide derivative, whether asingle compound or mixture of different conformers, have antifungalactivity as described herein. If desired, pure conformers may beisolated using techniques that are well-known in the art.

[0089] In some embodiments, the polyene macrolide derivatives of formula(I) are Amadori rearrangement products in which the reducingcarbohydrate reactant is selected from the group consisting of glucose,maltose, cellobiose, lactose, allose, and galactose, in either the α-,β-, D- or L-configurations, or mixtures thereof.

[0090] In the polyene macrolide amide derivatives of formula (I), theamide nitrogen has attached thereto and/or includes one or more of thesame or different water-solubility increasing substituents, designatedas R⁶ and/or R⁷. Water-solubility increasing substituents generallycomprise groups that have polar characteristics by virtue of includingone or more of the same or different heteroatoms. Such heteroatoms aretypically selected from the group consisting of O, N, NH and S, althoughother heteroatoms may be used.

[0091] In one embodiment, the water-solubility increasing substituentsare hydrocarbons such as linear and branched alkyls, cycloalkyls, arylsand arylalkyls that are substituted with one or more of the same ordifferent polar substituents. Typical polar substituents include, butare not limited to, —OH, —SH, ═O (oxo), ═S (thioxo), —NH₂, ═NH (imino),—C(═NH)—NH₂ (amidino), —NH—C(═NH)—NH₂ (guanidino), —C(O)H, —C(O)OH,—C(O)O⁻M⁺, —C(O)NH₂, —N₃, —CN, —X, —CX₃, etc., where each X isindependently a halogen, preferably F, Cl or Br and M⁺ represents amonovalent counter ion such as Na⁺, K⁺, etc. The polar substitutedalkyls, cycloalkyls, aryls and arylalkyls may be further optionallysubstituted with one or more of the same or different non-polarsubstituents, e.g., alkyls, cycloalkyls, aryls and arylalkyls, etc.;however care should be taken when selecting combinations of polar andnon-polar substituents to insure that the overall net character of thewater solubility increasing substituent is polar. Preferred polarsubstituted hydrocarbon water solubility increasing substituents arethose that include a plurality of polar substituents, such as, forexample (C₁-C₁₀) alkyls and (C₃-C₁₀) cycloalkyls that are substitutedwith a plurality of the same or different polar substituents. In oneembodiment, the plurality of polar substituents are, independently ofone another, selected from the group consisting of lower alkoxy,methoxy, hydroxy, amino, imino, amidino and guanidino groups.

[0092] In another embodiment, the heteroatom(s) is included in the chainof a hydrocarbon such that the water solubility-increasing substituentsare linear and branched heteroalkyls (e.g., ethers, thioethers,sulfonamides, alkylamines, etc.), cycloheteroalkyls (e.g.,imidazolidine, morpholine, piperazine, piperidine, pyrazolidine,pyrrolidine, quinuclidine, etc.), heteroaryls (e.g., chromane, chromene,furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran,isochromene, isoindole, isoindoline, isoquinoline, isothiazole,isoxazole, naphythyridine, phenanthroline, phthalazine, pteridine,purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine,pyrrole, pyrrolizine, quinazoline, quinoline, quinoxaline, thiophene,xanthene, etc.) and heteroarylalkyls (including, for example, compoundsof the formula (CH₂)_(m)R²², where m is an integer from 1 to 6 and R²²is a heteroaryl). One or more of the carbon atoms and/or heteroatoms(e.g., N, NH, etc.) of these heteroalkyl, cycloheteroalkyl, heteroaryland heteroarylalkyl groups may be further substituted with one or moreof the same or different polar or non-polar substituents, as describedabove. Again, the various polar and non-polar substituents should beselected such that the overall net character of the resultantwater-solubility increasing substituent will be polar. Preferredwater-solubility increasing substituents of this type are those whichinclude one or two of the same or different heteroatoms, and which areoptionally substituted at one or more of the carbon atoms orheteroatoms, as previously described. In one embodiment, the watersolubility increasing substituents are saturated or unsaturated 5-6membered rings that include one or two of the same or differentheteroatoms, preferably heteroatoms selected from the group consistingof O, N, NH and S. These heteroatom-containing rings may be optionallysubstituted with one or more of the same or different polar or non-polarsubstituents, as previously described. When such 5- or 6-membered ringsare substituted with non-polar substituents, the rings are usuallymono-substituted and the substituent is typically selected from thegroup consisting of (C₁-C₆) alkyl, (C₁-C₃₋₄) alkanyl, (C₅-C₆) aryl,phenyl, 6- to 9-membered arylalkyl and benzyl. The substituted orunsubstituted heteroatom-containing ring may be attached directly to theamide nitrogen or may be spaced away from the amide nitrogen via anamine, alkylamine or alkyldiyl spacer moiety, preferably a —NH—,(C₁-C₁₀) alkyldiyl, (C₁-C₁₀) alkyleno or (C₁-C₆) alkano spacer moiety.

[0093] In still another embodiment, R⁶ and R⁷ are taken together suchthat the amide nitrogen to which they are bonded is included as a memberof a saturated or unsaturated ring which may optionally include one ormore of the same or different additional heteroatoms and/or which isoptionally further substituted at one or more of the ring carbon orheteroatoms with the same or different polar or non-polar substituents,as previously described. In one embodiment, such rings are 5- or6-membered rings. When such 5- or 6-membered rings are substituted withnon-polar substitutents, the rings are usually mono-substituted and thesubstituent is typically selected from the group consisting of (C₁-C₆)alkyl, (C₁-C₃₋₄) alkanyl, (C₅-C₆) aryl, phenyl, 6- to 9-memberedarlyalkyl and benzyl.

[0094] In embodiments of the compounds of structural formulae (I) and(II) which include only a single water-solubility increasing substituentat the amide nitrogen, the other amide nitrogen substituent may behydrogen or a non-polar substituent, as previously described. In oneembodiment, the non-polar substituent is a lower alkyl, (C₁-C₃₋₄) alkyl,lower alkanyl or (C₁-C₃₋₄) alkanyl

[0095] As will be appreciated from the above discussion, the compositionof the water-solubility increasing substituents is not critical tosuccess. Further non-limiting examples of water-solubility increasingsubstituents that are useful include polyhydroxylated alkyls, mono-,di-, oligo- and polysaccharides, polyalkylene glycols (e.g.,polyethylene glycol, polypropylene glycol, etc.) and polyalkyleneoxides.

[0096] In one specific embodiment, the polyene macrolide amidederivatives are compounds according to structural formula (I) in which:

[0097] N—R¹—C(O), CH₂—R² and R¹⁴ are as previously described forstructural formula (I);

[0098] either: (i) R⁶ is selected from the group consisting of hydrogen,(C₁-C₆) alkyl, (C₁-C₆) alkyl substituted with one or more of the same ordifferent R¹⁰ groups, —[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—NR¹⁵R¹⁶,—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—NR¹⁵R¹⁶—[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—R¹⁷and —NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—R¹⁷ and R⁷ is selected from thegroup consisting of (C₁-C₆) alkyl substituted with one or more of thesame or different R¹⁰ groups, —[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—NR¹⁵R¹⁶,—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—NR¹⁵R¹⁶,—[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—R¹⁷ and—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—R¹⁷; or (ii) R⁶ and R⁷, taken togetherwith the amide nitrogen atom to which they are bonded, form a 5- or6-membered saturated or unsaturated ring which optionally includes oneor more of the same or different additional heteroatoms selected fromthe group consisting of O, N, NH and S and/or which is optionallysubstituted at one or more ring carbon or heteroatoms with the same ordifferent substituents selected from the group consisting of R¹⁰,(C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰, (C₅-C₆) aryl, phenyl, 6-to 9-membered arylalkyl and benzyl;

[0099] each R¹⁰ is independently selected from the group consisting of—OH, ═O (oxo), —NH₂ (amino), ═NH (imino), —C(═NH)—NH₂ (amidino) and—NH—C(═NH)—NH₂ (guanidino);

[0100] either: (i) R¹⁵ and R¹⁶ are each independently selected from thegroup consisting of hydrogen, (C₁-C₆) alkyl and (C₁-C₆) alkylindependently substituted with one or more of the same or different R¹⁰groups; or (ii) R¹⁵ and R¹⁶, taken together with the nitrogen atom towhich they are bonded, form a 5- or 6-membered saturated or unsaturatedring which optionally includes one or more of the same or differentadditional heteroatoms selected from the group consisting of O, N, NHand S and/or which is optionally substituted at one or more ring carbonor heteroatoms with the same or different substituents selected from thegroup consisting of R¹⁰, (C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰,(C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl and benzyl;

[0101] R¹⁷ is a 5- or 6-membered saturated or unsaturated ring whichoptionally includes one or more of the same or different additionalheteroatoms selected from the group consisting of O, N, NH and S and/orwhich is optionally substituted at one or more ring carbon orheteroatoms with the same or different substituents selected from thegroup consisting of O, N, NH and S and/or which is optionallysubstituted at one or more ring carbon or heteroatoms with the same ordifferent substituents selected from the group consisting of R¹⁰,(C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰, (C₅-C₆) aryl, phenyl, 6-to 9-membered arylalkyl and benzyl;

[0102] each n is independently an integer from 1 to 6; and

[0103] each p is independently an integer from 0 to 6.

[0104] In one specific embodiment of the above-described compounds, R¹⁵and R¹⁶ are defined according to alternative (ii) and form a saturatedring which optionally includes additional heteroatoms and which isoptionally substituted, as described above. In another specificembodiment, R¹⁷ is a 5- to 6-membered heteroaryl ring, optionallysubstituted as described above.

[0105] In another embodiment, the polyene amide derivatives ofstructural formula (I) and/or any of the above-described specificembodiments thereof, where applicable, have one or more featuresselected from the group consisting of:

[0106] N—R¹—C(O) is a polyene backbone derived from AmB or nystatin;

[0107] CH₂—R¹ is a mono-, di- or oligosaccharide;

[0108] R⁶ is hydrogen; and/or

[0109] R¹⁴ is hydrogen.

[0110] In still another embodiment, the polyene macrolide derivatives ofthe invention are compounds according to structural formula (I) inwhich:

[0111] R⁶ is hydrogen;

[0112] R⁷ is selected from the group consisting of —NH—NR¹⁵R¹⁶,—(CH₂)_(n)—NR¹⁵R¹⁶, —(CH₂)_(n)—R¹⁷ and (C₁-C₆) alkyl substituted withone or more amino or hydroxyl groups;

[0113] R¹⁵ and R¹⁶, taken together with the nitrogen atom to which theyare bonded, form a 5- or 6-membered saturated or unsaturated ring whichoptionally includes one or more additional heteroatoms selected from thegroup consisting of O, S, N and NH and/or which is optionallysubstituted at one or more ring carbon or heteroatoms with the same ordifferent R¹⁰, (C₁-C₆) alkyl, (C₁-C₆) alkoxy, (C₅-C₆) aryl, phenyl, 6-to 9-membered arylalkyl or benzyl groups;

[0114] R¹⁷ is a 5- or 6-membered heteroaryl which is optionallysubstituted with one or more of the same or different (C₁-C₆) alkyl,(C₁-C₆) alkoxy, (C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl orbenzyl groups; and

[0115] all other variables are as defined for structural formula (I).

[0116] In still another embodiment, the polyene macrolide amidederivatives of the invention are compounds according to structuralformula (I) in which:

[0117] R⁶ and R⁷, taken together with the nitrogen atom to which theyare bonded, form a 5- to 6-membered cycloheteroalkyl ring optionallysubstituted with one or more, preferably one, substituent selected fromthe group consisting of (C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰,(C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl and benzyl;

[0118] R¹⁰ is amino or hydroxy; and

[0119] all other variables are as defined for structural formula (I). Inone embodiment, the cycloheteroalkyl ring is a morpholine or apiperazine ring. When substituted, the piperazine ring is preferablysubstituted at the N-position.

[0120] Exemplary water-solubility increasing substituents useful in thecontext of the polyene macrolide amide derivatives according tostructural formula (I) are illustrated with reference to compounds(100)-(122) and (127)-(147), infra. Additional exemplary watersolubility increasing substituents are described in U.S. Pat. No.5,296,597 to Bruzzese et al., (see especially Cols. 6-10 and Table 1),U.S. Pat. No. 5,298,495 to Bruzzese et al., (see especially Tables 1 and2) and Bruzzese et al., 1996, Eur. J. Med. Chem. 31:965-972 (seeespecially Table I), the disclosures of which are incorporated herein byreference.

[0121] Exemplary polyene macrolide amide derivatives according tostructural formula (I) in which carbohydrate CH₂—R² is an Amadorirearrangement product using α-D-glucose as the reducing carbohydrateinclude the following compounds: (100)

(101)

(102)

(103)

(104)

(105)

(106)

(107)

(127)

(128)

(129)

(130)

(131)

[0122] Also exemplified are the corresponding nystatin derivatives ofCompounds (100) through (107) and (127) through (131).

[0123] Exemplary polyene macrolide amide derivatives according tostructural formula (I) in which carbohydrate CH₂—R² is an Amadorirearrangement product using D-galactose as the reducing carbohydrateinclude the following compounds: (108)

(109)

(110)

(111)

(112)

(113)

(114)

(115)

(132)

(133)

(143)

(135)

(136)

(137)

[0124] Also exemplified are the corresponding nystatin derivatives ofCompounds (108) through (115) and (132) through (137).

[0125] Exemplary polyene macrolide amide derivatives according tostructural formula (I) in which carbohydrate CH₂—R² is an Amadorirearrangement product using D-maltose as the reducing carbohydrateinclude the following compounds: (116)

(117)

(118)

(119)

(120)

(121)

(122)

(138)

(139)

(140)

(141)

(142)

(143)

(144)

[0126] Also exemplified are the corresponding nystatin derivatives ofCompounds (141) through (144).

[0127] Exemplary polyene macrolide amide derivatives according tostructural formula (I) in which carbohydrate CH₂—R² is an Amadorirearrangement product using α-D-lactose as the reducing carbohydrateinclude the following compounds: (145)

(146)

(147)

[0128] Also exemplified are the corresponding nystatin derivatives ofCompounds (144) through (147).

[0129] In polyene macrolide amide derivatives according to structuralformula (II), R⁴ and R¹⁴ are both preferably hydrogen or one is hydrogenand the other is (C₁-C₃) alkanyl, R³ is hydrogen and/or R⁵ is amonosaccharide.

[0130] Exemplary polyene macrolide amide derivatives according tostructural formula (II) include the following compounds: (123)

(124)

(125)

(126)

[0131] Also exemplified are the corresponding nystatin derivatives ofCompounds (123) through (126).

[0132] Those of skill in the art will appreciate that many of thecompounds encompassed by formulae (I) and (II), as well as the compoundspecies specifically described herein, may exhibit the phenomena oftautomerism, conformational isomerism, geometric isomerism and/or stereoisomerism. As the formula drawings within the specification and claimscan represent only one of the possible tautomeric, conformationalisomeric, enantiomeric or geometric isomeric forms, it should beunderstood that the invention encompasses any tautomeric, conformationalisomeric, enantiomeric and/or geometric isomeric forms of the compoundshaving one or more of the utilities described herein, as well asmixtures of these various different forms.

[0133] Moreover, in many of the compounds, the polyene backboneN—R¹—C(O) is illustrated with the stereochemistry of many of the chiralcenters specified. The specific structures depicted are those that havebeen reported in the literature for the involved polyene backbones, andare not intended as limiting. Thus, it will be understood that theillustrated structures are intended merely as a short-hand way ofrepresenting the actual compounds, and to the extent it may be found ata later date that these structural representations are incorrect, theyare not intended to be limiting in any way.

[0134] The polyene macrolide amide derivatives of formulae (I) and (II)may be synthesized according to well-known methods using well-knownchemistries. In one embodiment, the polyene macrolide amide derivativesof formula (I) may be synthesized according to Scheme (I), illustratedbelow in which R⁶ and R¹⁴ are each hydrogen:

[0135] In Scheme (I), N—R¹—C(O), —CH₂—R² and R⁷ are as previouslydefined for structural formula (I). According to Scheme (I), a parentpolyene macrolide 10 having an exocyclic carboxyl group and an aminofunctionality is reacted with a reducing carbohydrate 13, for exampleone of the reducing carbohydrates listed in TABLE 1, supra, underAmadori rearrangement conditions to yield glycosylated Amadori product12. The Amadori rearrangement reaction is described in detail inAmadori, 1955, Adv. Carbohydr. Chem. 10:169 and Hodge & Fisher, supra,both of which are incorporated herein by reference. Amadori product 12is then converted to the corresponding amide 14 by reaction with amine11 (preferably in the free base form) using standard methods that arewell known in the art (see, e.g., Bruzzese, 1996, Eur. J. Med. Chem.31:965-992) or the methods described below. Compounds according tostructural formula (I) having different water solubility increasingsubstituents at the amide nitrogen may be synthesized in an analogousmanner by reacting Amadori product 12 with an appropriate amine 11. Ifnecessary, any reactive substituents on amine 11 and/or carbohydrateCH₂—R² may be protected using well-known protecting groups andchemistries. The actual protecting group selected will depend upon,among other factors, the identity of the reactive substituent, and willbe apparent to those of skill in the art. Non-limiting examples ofprotecting groups suitable for a wide variety of reactive groups, aswell as conditions for their attachment and removal, can be found inGreene & Wats, Protective Groups in Organic Synthesis, 3^(rd) Edition,John Wiley & Sons, Inc., NY (1999), which is incorporated by reference.

[0136] Alternatively, polyene macrolide amide derivatives of formula (I)may be synthesized according to Scheme (II) below, in which R⁶ and R¹⁴are each hydrogen:

[0137] In Scheme (II), N—R¹—C(O), CH—R² and R⁷ areas previously definedfor structural formula (I). According to Scheme (II), parent polyenemacrolide 10 is reacted with amine 11 (preferably in the free base form)under standard conditions to yield amidated intermediate 16. Amidatedintermediate 16 is then reacted with reducing carbohydrate 13 underAmadori rearrangement conditions to yield polyene amide derivative 14.If necessary, any reactive substituents on amine 11 and/or reducingcarbohydrate 13 may be protected, as previously described for Scheme(I). Compounds according to structural formula (I) including two watersolubility increasing substituents at the amide nitrogen, e.g. compoundsof formula (I) in which R⁶ and R⁷ are each other than hydrogen, may besynthesized by routine modification of the illustrated methods byselecting the appropriate amine 11. Compounds according to structuralformula (I) in which the amide nitrogen is included in a ring (e.g.,compound 106) may be synthesized by routine modification of theillustrated methods by selecting the appropriate amine 11.

[0138] It has been discovered that the amidation steps illustrated inSchemes (I) and (II) may be advantageously carried out using chemistriesand reagents commonly employed in peptide chemistry for the formation ofamide bonds. In one embodiment, the amidation steps may be carried outusing an uronium salt, such as, for example,O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(“HBTU”); O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (“HATU”); O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-bis(tetramethylene)uronium hexafluorophosphate (“HAPyU”);O-(benzotriazol-1-yl)-N,N, N′,N′-bis(pentamethylene)uroniumhexafluorophosphate (“HBPipU”);O-(benzotriazol-1-yl)-N,N,N′,N′-bis(tetramethylene)uroniumhexafluorophosphate (“HBPyU”); O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (“TBTU”);O-(benzotriazol-1-yl)-N,N,N′,N′-bis(pentamethylene)uroniumtetrafluoroborate; O-[4-oxo01,2,3-benzotriazin-3(4H)-yl]-N,N,N′,N′-tetramethyluronium tetrafluoroborate (“TDBTU”);O-(1,2-dihydro-2-oxo-1-puridyl)-N,N,N′,N′-tetramethyluroniumtetrafluoroborate (“TPTU”);O-(N-succinimidyl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate(“TSTU”); andO-(5-norborene-2,3-dicarboximido)-N,N,N′,N′-tetramethyluroniumtetrafluoroborate (“TNTU”). The reactions may be carried out asdescribed, for example, in Dourtuglou, Synthesis, 572 (1984), thedisclosure of which is incorporated herein by reference. Briefly, thereactions are typically carried out in the presence of a base, which ismost usually an unhindered or hindered amine base such as, for example,diisopropylethyl amine (“DIEA”), N-methylmorpholine or triethylamine,and the desired amine 11 in a polar organic solvent, usually a dipolarorganic solvents such as, for example, dimethyl sulfoxide (“DMSO”),dimethylacetamide (“DMA”) or dimethylformamide (“DMF”), at a temperaturein the range of about −10° C. to 30° C. The ratios of carboxylic acid 10or 12, uronium salt, base and amine 11 are typically in the range of1:1:1-3:excess or 1:1.5:1.5-5:1.5, although other ratios may be used.

[0139] In another embodiment, the amidation steps may be carried outusing a phosphonium salt, such as, for example,benzotriazole-1-yloxy-tris(dimethylamino) phosphoniumhexafluorophosphate (“BOP”); bromo-tris-pyrrolidino-phosphoniumhexalfluorophosphate (“PyBroP”); andbenzotriazole-1-yloxy-tripyrrolodino phosphonium hexafluorophosphate(“PyBOP”). The reactions may be carried out as described above for theuronium salts.

[0140] It has also been discovered that polyene amide derivatives offormula (I) may be synthesized in a one-pot reaction with high yield.The one-pot reaction is illustrated in Scheme (III) below, in which R⁶and R¹⁴ are each hydrogen:

[0141] Substituents N—R¹—C(O), CH₂—R² and R⁷ are as defined forstructural formula (I). According to Scheme (III), parent polyenemacrolide 10 (1 equiv) and reducing carbohydrate 13 (about 1.2 to about1.5 equiv.) are dissolved in a suitable organic solvent, typically adipolar solvent such as DMF, and stirred at a temperature in the rangeof about 30° C. to about 55° C. for about 5 to 72 hr. The reaction maybe followed by TLC, HPLC or other routing methods to insure that thefirst step of the reaction (Amadori rearrangement) has gone to asatisfactory level of completion. The reaction mixture is then cooled,typically to a temperature in the range of about 10° C. to about 5° C.In one embodiment, once cooled, a uronium salt (about 1.5 to about 3equiv.) or phosphonium salt (about 1.5 to about 3 equiv.) couplingreagent and an appropriate base, for example, DIEA (about ______ toabout ______ equiv.) is added, stirred for a few minutes (e.g., fromabout 5 to about 30 min.), and amine 11 (about 1.5 to about 5 equiv.) isadded. In other embodiments, the amidation reagents are added in otherorders. For example, the amidation reagents may be added simultaneously,or the amine 11 may be added first followed by the coupling reagent andDIEA, etc. The order of reagent addition for the amidation step is notcritical to success. The reaction mixture is then warmed, typically to atemperature in the range of about 20° C. to about 30° C., and thereaction allowed to proceed to completion, which generally takes about0.5 to about 18 hr. The polyene macrolide amide derivative may berecovered from the reaction mixture using standard techniques, such asprecipitation or chromatography.

[0142] Polyene macrolide amide derivatives of formula (I) in whichsubstituent R¹⁴ is an alkyl may be synthesized in a variety of differentways, illustrated in Scheme (IV), infra:

[0143] In Scheme IV, substituents N—R¹—C(O), CH₂—R², R⁷ are as definedfor structural formula (I), R⁶ is hydrogen, R¹⁴ is alkyl and X is ahalogen, such as Cl, Br or I. According to Scheme IV, in one method,parent polyene macrolide 10 is first mono-alkylated with alkylatingreagent 20 according to standard techniques to yield alkylated parentpolyene macrolide 22. Although alkylating reagents 20 is illustrated asa halogenated alkyl, skilled artisans will recognize that virtually anystandard alkylating reagents may be used. Alkylated parent macrolide 22may then be converted to alkylated polyene macrolide amide derivative 28according to the methods described in Schemes I, I or III.

[0144] In another method, glycosylated Amadori product 12 is alkylatedusing standard alkylation techniques to yield alkylated Amadori product26, which is then amidated according to any of the previously describedmethods to yield alkylated polyene macrolide amide derivative 28. Instill another method, amidated polyene macrolide 16 is alkylated usingstandard alkylation techniques to yield alkylated amidated polyenemacrolide 24, which is then reacted with carbohydrate 13 under Amadorirearrangement conditions to yield alkylated polyene macrolide amidederivative 28.

[0145] Polyene macrolide amide derivatives of formula (II) may besynthesized by routine modification of the above-described methods.Mono- and dialkylated derivatives may be synthesized by first alkylatingthe parent polyene macrolide 10, followed by amidation, or,alternatively, by first amidating parent polyene macrolide 10, followedby mono- or dialkylation according to standard techniques.

[0146] Parent polyene macrolide 10 may be obtained commercially or maybe isolated or synthesized according to well-known methods. Methods ofsynthesizing a variety of parent polyene macrolides 10 are described inBeau, “Polyene Macrolides: Stereostructural Elucidation and SyntheticStudies of a Few Members,” In: Recent Progress in the Chemical Synthesisof Antibiotics, pp. 135-182, Springer-Verlag, Berlin (1990), as well asthe references cited therein. These methods may be routinely adapted tosynthesize a wide variety of parent polyene macrolides 10. Methods ofisolating parent polyene macrolides 10 as natural products arewell-known in the art.

[0147] In addition to the above-described methods, methods forconverting parent polyene macrolide 10 or glycosylated Amadori product12 to amides are well-known in the art. Exemplary methods are describedin U.S. Pat. No. 4,783,527. Specific methods of synthesizing amides ofAmB are described in Czerwinski et al., 1990, J. Antibiot. 43(6):680-683and Jarzebski et al., 1982, J. Antibiot. 35(2):220-229. Specific methodsof synthesizing amides of patricins are described in U.S. Pat. No.5,298,495, U.S. Pat. No. 5,296,597 and Bruzzese et al., 1996, J. Med.Chem. 31:965-972. Any of these methods may be routinely adapted tosynthesize the full range of mono- and di-substituted polyene macrolideamide derivatives of the invention. All of the above-listed patents andreferences, as well as the various patents and references cited therein,are incorporated herein by reference.

[0148] In all of the illustrated schemes, the Amadori rearrangementreaction proceeds in two steps. Referring to Scheme (I), the formationof glycosylated Amadori product 12 proceeds in two steps. In the firststep, a glycosylamine (not shown) is formed by condensation of theprimary amino group of 10 with the anomeric carbon of reducingcarbohydrate 13. In the second step, the glycosylamine is rearranged inan acidic medium to form the glycosylated Amadori product 12. Thus, areducing carbohydrate 13 reactant should be selected that, after theAmadori rearrangement, will yield the desired carbohydrate residueCH₂—R².

[0149] The principles of the Amadori rearrangement are well-known andhave been briefly illustrated supra. Thus, choosing an appropriatereducing carbohydrate 13 will be apparent to those of skill in the art.Specific exemplary reducing carbohydrates 13 are provided in TABLE 1.Additional guidance can be found in Amadori, 1955, Adv. Carbohydr. Chem.10:169, Hodge & Fisher, supra, (and the references cited therein) andU.S. Pat. No. 5,314,999, all of which are incorporated herein byreference.

[0150] The Amadori rearrangement is acid-catalyzed. Since many of thedescribed rearrangement reactions involve polyene macrolides having anacidic carboxyl substituent, these carboxyl-containing polyenemacrolides can “self-catalyze” the rearrangement. Such “self-catalyzed”Amadori rearrangement reactions (e.g., the reactions of Scheme I) may becarried out in virtually any solvent system known in the art to beuseful for performing Amadori rearrangement reactions in which theparent polyene macrolide is stable, inlcuding the anhydrous solventsystems described in the literature (see, e.g. Hodge & Fischer, supra).For AmB; which is not very stable in acids and bases, overly acidic andbasic solvent systems, such as, for example, glacial acetic acid, shouldbe avoided.

[0151] Unlike Scheme (I) and many of the Amadori rearrangements reportedin the literature, in the method of Scheme (II), derivatives 16 do nothave a free carboxyl group. Rather, the exocyclic carboxyl of parentpolyene macrolide 10 has been amidated. Thus, derivatives 16 may notefficiently “self-catalyze” the Amadori rearrangement. As a consequence,it has been discovered that it is preferable to conduct the Amadorirearrangement reaction between amidated derivative 16 and reducingcarbohydrate 13 in the presence of water. While the reaction willproceed under anhydrous conditions, significantly better yields areobtained under non-anhydrous conditions. A variety of non-anhydroussolvent systems may be used for Scheme (II). Typically, the solventsystem should comprise about 1% (v/v) to 5% (v/v) water. The protondonor may be the solvent system or it may be an added compound; asdescribed in Hodge & Fisher, supra. Any of the solvent systems describedin the literature may be adapted for use as described herein. Exemplarysolvent systems that may be readily adapted to the principles taughtherein are described in Hodge & Fisher, 1963, supra. Specific solventsystems are provided in the Examples section, infra.

[0152] When synthesizing polyene macrolide amide derivatives accordingto Scheme (I), it has been discovered that using N,N′-dimethylpropyleneurea (“DMPU”) as the solvent for the Amadori rearrangement step yieldsbetter results as compared with other solvents, such as DMF. Forexample, when DMF is used as the solvent, a small but measurablequantity of a side product is produced. By comparison, less of this sideproduct is produced when the reaction is carried out in DMPU.

[0153] Moreover, the literature reports that the Amadori rearrangementmay be successfully performed with a 1:1 molar ratio of polyenemacrolide:reducing carbohydrate, e.g., compounds 10 and 13 in Scheme(I). However, it has been discovered that using a molar ratio in therange of about 1:1.1 yields better results. It has also been discoveredthat adding the reducing carbohydrate in three equal aliquots at equalintervals of about 1.25 hr to about 1.5 hr, yields better results. Thus,while the Amadori rearrangement reactions illustrated in the aboveschemes may be performed with 1 equivalent of reducing carbohydrate 13,using about 1.1 total equivalents of reducing carbohydrate 13, added inthree equal aliquots of about 0.367 equiv, added at intervals of about1.25 hr to about 1.5 hr, is preferred.

[0154] The polyene macrolide amide derivatives of the invention exhibitsignificant antifungal activity, typically having minimum inhibitoryconcentrations (MICs) of about 8 μg/mL or less against C. albicans instandard in vitro assays. Generally, active polyene macrolide amidederivatives of the invention are identified using in vitro screeningassays that are well-known in the art. Specific in vitro screeningassays that can be used to assess activity are provided in the Examplessection.

[0155] Alternatively, the polyene macrolide amide derivatives of theinvention may be assessed for antifungal activity using in vivo models.Again, such models are well-known in the art. Other assays as are wellknown in the art, or that will become apparent to those having skill inthe art upon review of this disclosure, may also be used to identifyactive polyene macrolide amide derivatives of the invention.

[0156] Generally, active polyene macrolide amide derivatives of theinvention will exhibit minimum inhibitory concentrations (MICs) of lessthan about 64 μg/mL, usually less than about 32 μg/mL, preferably lessthan about 16 μg/mL and most preferably less than about 8 μg/mL againstCandida albicans using standard methods. Of course, compounds havingMICs on the low end of these ranges, or even lower, are preferred.

[0157] All of the derivatives of the invention may be used topically orsystemically, as will be described in more detail, below. For in vivoapplications, such as for systemic administration and/or for use intreating or preventing systemic infections, derivatives that exhibitsignificant antifungal activity (i.e., less than 4 μg/mL), higherwater-solubility than AmB (at approx. neutral pH) and low toxicity arepreferred. Generally, derivatives which exhibit an ED₅₀ of ≦20 instandard mouse kidney bioburden assays, such as the 5 day and 7 daymouse bioburden assays described in the Examples section, are suitablefor in vivo applications. Toxicity is less of a concern for topicaladministration and applications, as is water solubility.

[0158] The polyene macrolide amide derivatives of the present inventionhave significant advantages over currently available polyene macrolideantifungals. Specifically, the polyene macrolide amide derivatives ofthe present invention show excellent water solubility, low toxicity, andeffective therapeutic potency. Moreover, both classes of derivatives,i.e., compounds according to formulae (I) and (II), exhibit antifungalactivity comparable to AmB in both in vitro and in vivo assays, withlower acute toxicity.

[0159] The polyene macrolide amide derivatives according to theinvention can be used in a wide variety of applications to inhibit thegrowth of or kill fungi. For example, the polyene macrolide amidederivatives can be used as disinfectants or as preservatives formaterials such as foodstuffs, cosmetics, medicaments and othernutrient-containing materials.

[0160] For use as a disinfectant or preservative, the polyene macrolideamide derivatives can be added to the desired material singly, asmixtures of several polyene macrolide amide derivatives, or incombination with other antifungal and/or antimicrobial agents. Thepolyene macrolide amide derivatives may be supplied as the compound perse or may be in admixture with a variety of carriers, diluents orexcipients as are well known in the art.

[0161] When used to treat or prevent fungal infections the polyenemacrolide amide derivatives of the invention can be administered orapplied singly, as mixtures of two or more polyene macrolide amidederivatives, in combination with other antifungal, antibiotic orantimicrobial agents or in combination with other pharmaceuticallyactive agents. The polyene macrolide amide derivatives can beadministered or applied per se or as pharmaceutical compositions. Thespecific pharmaceutical formulation will depend upon the desired mode ofadministration, and will be apparent to those having skill in the art.Numerous compositions for the topical or systemic administration ofpolyene macrolides are described in the literature. Any of thesecompositions may be formulated with the polyene macrolide amidederivatives of the invention.

[0162] Pharmaceutical compositions comprising the polyene macrolideamide derivatives of the invention may be manufactured by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or lyophilizing processes.Pharmaceutical compositions may be formulated in conventional mannerusing one or more physiologically acceptable carriers, diluents,excipients or auxiliaries which facilitate processing of the activepolyene macrolide amide derivatives into preparations which can be usedpharmaceutically. Proper formulation is dependent upon the route ofadministration chosen.

[0163] For topical administration the polyene macrolide amidederivatives of the invention may be formulated as solutions, gels,ointments, creams, suspensions, etc. as are well-known in the art.

[0164] Systemic formulations include those designed for administrationby injection, e.g. subcutaneous, intravenous, intramuscular, intrathecalor intraperitoneal injection, as well as those designed for transdermal,transmucosal, oral or pulmonary administration.

[0165] For injection, the polyene macrolide amide derivatives of theinvention may be formulated in aqueous solutions, preferably inphysiologically compatible buffers such as Hanks's solution, Ringer'ssolution, or physiological saline buffer. The solution may containformulatory agents such as suspending, stabilizing and/or dispersingagents.

[0166] Alternatively, the polyene macrolide amide derivatives may be inpowder form for constitution with a suitable vehicle, e.g., sterilepyrogen-free water, before use.

[0167] For transmucosal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art.

[0168] For oral administration, the polyene macrolide amide derivativescan be readily formulated by combining them with pharmaceuticallyacceptable carriers well known in the art. Such carriers enable thecompounds of the invention to be formulated as tablets, pills, dragees,capsules, liquids, gels, syrups, slurries, suspensions and the like, fororal ingestion by a patient to be treated. For oral solid formulationssuch as, for example, powders, capsules and tablets, suitable excipientsinclude fillers such as sugars, such as lactose, sucrose, mannitol andsorbitol; cellulose preparations such as maize starch, wheat starch,rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP); granulating agents; and binding agents. Ifdesired, disintegrating agents may be added, such as the cross-linkedpolyvinylpyrrolidone, agar, or alginic acid or a salt thereof such assodium alginate.

[0169] If desired, solid dosage forms may be sugar-coated orenteric-coated using standard techniques.

[0170] For oral liquid preparations such as, for example, suspensions,elixirs and solutions, suitable carriers, excipients or diluents includewater, glycols, oils, alcohols, etc. Additionally, flavoring agents,preservatives, coloring agents and the like may be added.

[0171] For buccal administration, the compositions may take the form oftablets, lozenges, etc. formulated in conventional manner.

[0172] For administration by inhalation, the compounds for use accordingto the present invention are conveniently delivered in the form of anaerosol spray from pressurized packs or a nebulizer, with the use of asuitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitmay be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, e.g., gelatin for use in an inhaler orinsufflator may be formulated containing a powder mix of the compoundand a suitable powder base such as lactose or starch.

[0173] The compounds may also be formulated in rectal or vaginalcompositions such as suppositories or retention enemas, e.g., containingconventional suppository bases such as cocoa butter or other glycerides.

[0174] In addition to the formulations described previously, the polyenemacrolide amide derivatives may also be formulated as a depotpreparation. Such long acting formulations may be administered byimplantation (for example subcutaneously or intramuscularly) or byintramuscular injection. Thus, for example, the compounds may beformulated with suitable polymeric or hydrophobic materials (for exampleas an emulsion in an acceptable oil) or ion exchange resins, or, assparingly soluble derivatives, for example, as a sparingly soluble salt.

[0175] Alternatively, other pharmaceutical delivery systems may beemployed. Liposomes and emulsions are well known examples of deliveryvehicles that may be used to deliver the polyene macrolide amidederivatives of the invention. Certain organic solvents such asdimethylsulfoxide also may be employed, although usually at the cost ofgreater toxicity. Additionally, the polyene macrolide amide derivativesmay be delivered using a sustained-release system, such as semipermeablematrices of solid polymers containing the therapeutic agent. Various ofsustained-release materials have been established and are well known bythose skilled in the art. Sustained-release capsules may, depending ontheir chemical nature, release the compounds for a few weeks up to over100 days.

[0176] As certain substituents on the polyene macrolide amidederivatives of the invention may be acidic or basic, the derivatives maybe included in any of the above-described formulations as the freeacids, the free bases or as pharmaceutically acceptable salts.Pharmaceutically acceptable salts are those salts which retainsubstantially the antifungal activity of the free acids or bases andwhich are prepared by reaction with bases or acids, respectively.Pharmaceutical salts tend to be more soluble in aqueous and other proticsolvents than are the corresponding free base or acid forms. Someexamples of pharmaceutically acceptable salts include: (1) acid additionsalts, formed with inorganic acids such as hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike; or formed with organic acids such as amino acids (e.g., asparticacid, glutamic acid, asparagine, glutamine, lysine, ornithine) aceticacid, propionic acid, hexanoic acid, cyclopentanepropionic acid,glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid,malic acid, maleic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelicacid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the amide derivative iseither replaced by a metal ion, e.g., an alkali metal ion, an alkalineearth ion, or an aluminum ion; or coordinates with an organic base suchas ethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like. In one embodiment, pharmaceuticallyacceptable salts are formed with aspartic acid, glutamic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid and mandelic acid. Inanother embodiment, pharmaceutically acceptable salts are formed withaspartic acid, glutamic acid, and fumaric acid.

[0177] The polyene macrolide amide derivatives of the invention, orcompositions thereof, will generally be used in an amount effective toachieve the intended purpose. Of course, it is to be understood that theamount used will depend on the particular application.

[0178] For example, for use as a disinfectant or preservative, anantifungally effective amount of a polyene macrolide derivative, orcomposition thereof, is applied or added to the material to bedisinfected or preserved. By antifungally effective amount is meant anamount of polyene macrolide derivative or composition that inhibits thegrowth of, or is lethal to, a target fungi. While the actual amount willdepend on a particular target fungi and application, for use as adisinfectant or preservative the polyene macrolide amide derivatives, orcompositions thereof, are usually added or applied to the material to bedisinfected or preserved in relatively low amounts. Typically, thepolyene macrolide derivative comprises less than about 5% by weight ofthe disinfectant solution or material to be preserved, preferably lessthan about 1% by weight and more preferably less than about 0.1% byweight. An ordinarily skilled artisan will be able to determineantifungally effective amounts of particular polyene macrolide amidederivatives for particular applications without undue experimentationusing, for example, the in vitro assays provided in the examples.

[0179] For use to treat or prevent fungal infections, the polyenemacrolide amide derivatives of the invention, or compositions thereof,are administered or applied in a therapeutically effective amount. Bytherapeutically effective amount is meant an amount effective toameliorate the symptoms of, or ameliorate, treat or prevent fungalinfections. Determination of a therapeutically effective amount is wellwithin the capabilities of those skilled in the art, especially in lightof the detailed disclosure provided herein.

[0180] As in the case of disinfectants and preservatives, for topicaladministration to treat or prevent fungal infections, a therapeuticallyeffective dose can be determined using, for example, the in vitro assaysprovided in the examples. The treatment may be applied while theinfection is visible, or even when it is not visible. An ordinarilyskilled artisan will be able to determine therapeutically effectiveamounts to treat topical infections without undue experimentation.

[0181] For systemic administration, a therapeutically effective dose canbe estimated initially from in vitro assays. For example, a dose can beformulated in animal models to achieve a circulating polyene macrolidederivative concentration range that includes the MIC as determined incell culture. Such information can be used to more accurately determineuseful doses in humans.

[0182] Initial dosages can also be estimated from in vivo data, e.g.,animal models, using techniques that are well known in the art. Onehaving ordinary skill in the art can readily optimize administration tohumans based on animal data.

[0183] Alternatively, initial dosages can be determined from the dosagesadministered of known polyene macrolides (e.g., AmB) by comparing theMIC of the specific polyene macrolide derivative with that of a knownpolyene macrolide, and adjusting the initial dosages accordingly. Theoptimal dosage may be obtained from these initial values by routineoptimization.

[0184] Dosage amount and interval may be adjusted individually toprovide plasma levels of the active polyene macrolide derivative whichare sufficient to maintain therapeutic effect. Usual patient dosages foradministration by injection range from about 0.1 to 5 mg/kg/day,preferably from about 0.5 to 1 mg/kg/day. Therapeutically effectiveserum levels may be achieved by administering a single daily dose ormultiple doses each day.

[0185] As the polyene macrolide amide derivatives of the inventionexhibit lower toxicity than AmB they can be administered in a mannersimilar to AmB. Typical dosages and routes of administration used forAmB are well-known (see, e.g., Goodman and Gilman's The PharmacologicalBasis of Therapeutics, 8^(th) Edition, 1990, Pergamon Press Inc., pp.1165-1168, incorporated herein by reference).

[0186] For example, a small test dose (1 mg compound dissolved in 20 mlof 5% dextrose solution) may be administered intravenously over 20-30min. The temperature, pulse, respiratory rate and blood pressure of thepatient may be recorded every 30 min. for 4 hrs. A patient with asevere, rapidly progressing fungal infection, good cardiopulmonaryfunction and a mild reaction to the test dose can immediately receive0.3 mg/kg compound intravenously over a period of 2 to 4 hrs (see, e.g.,Beunet, 1990 “Antifungal Agents,” In: Principles and Practice ofInfectious Diseases, 3^(rd) Ed., Churchill Livingstone, Inc., New York,pp. 361-370). If the patient has a severe reaction to the test dose orcardiopulmonary impairment, a smaller dose may be recommended, forexample, 0.1 mg/kg or 5-10 mg. This dose may be increased by 5-10 mg perday. In severe or fulminant infections, dosage should be escalatedrapidly until the patient is receiving 0.5 to 1.0 mg/kg daily.

[0187] Incremental doses can be given every 6 to 8 hrs if reactions in afragile patient make immediate advancement to full dosage inadvisable.For example, a severe reaction to a 1 mg test dose could be followed by5, 15 and 25 mg given at 8 hr intervals, followed by 40 mg 24 hrs later.The recommended maintenance dose for most deep mycoses is 0.4 to 0.6mg/kg/day, infused over 2-4 hrs.

[0188] In cases of local administration or selective uptake, theeffective local concentration of polyene macrolide derivative may not berelated to plasma concentration. One having skill in the art will beable to optimize therapeutically effective local dosages without undueexperimentation.

[0189] The amount of polyene macrolide amide derivative administeredwill, of course, be dependent on, among other factors, the subject beingtreated, the subject's weight, the severity of the affliction, themanner of administration and the judgment of the prescribing physician.

[0190] The antifungal therapy may be repeated intermittently whileinfections are detectable, or even when they are not detectable. Thetherapy may be provided alone or in combination with other drugs, suchas for example other antifungals, antibiotics or antimicrobials, orother polyene macrolide amide derivatives of the invention.

[0191] Preferably, a therapeutically effective dose of the polyenemacrolide amide derivatives described herein will provide therapeuticbenefit without causing substantial toxicity. Toxicity of the polyenemacrolide amide derivatives can be determined using standardpharmaceutical procedures in cell cultures or experimental animals,e.g., by determining the LD₅₀ (the dose lethal to 50% of the population)or the LD₁₀₀ (the dose lethal to 100% of the population). The dose ratiobetween toxic and therapeutic effect is the therapeutic index. Polyenemacrolide amide derivatives which exhibit high therapeutic indices arepreferred. The data obtained from these cell culture assays and animalstudies can be used in formulating a dosage range that is not toxic foruse in humans. The dosage of the polyene macrolide amide derivativesdescribed herein lies preferably within a range of circulatingconcentrations that include the effective dose with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized. The exactformulation, route of administration and dosage can be chosen by theindividual physician in view of the patient's condition. See, e.g.,Fingl et al., 1975, In: The Pharmacological Basis of Therapeutics, Ch.1,p.1.

[0192] The invention having been described, the following examples arepresented to illustrate, rather than to limit, the scope of theinvention.

5. EXAMPLE Preparation of Amide Derivatives of Amphotericin B

[0193] This example demonstrates the preparation of various amidederivatives of amphotericin B (“AmB”). AmB was purchased from BiosourcePharm, Spring Valley, N.Y. Amines were purchased from Aldrich(Milwaukee, Wis.) or Fluka (Milwaukee, Wis.). All of the compoundssynthesized are illustrated in TABLE 2, which is presented at the end ofthe example. In TABLE 2, the amine reducing carbohydrate used in theamidation and Amadori rearrangement, respectively, are listed. All ofthe compounds described in TABLE 2, including those that were notspecifically synthesized (as demonstrated by the absence of a compoundnumber in the particular table cell) are expected to be active and canbe synthesized according to the methods described in this section.

[0194] The polyene macrolide amide derivatives are sensitive to light,and should therefore be protected from light during preparation andstorage. Storage in amber vials is recommended.

[0195] 5.1 LCMS and HPLC Analyses

[0196] The various reactions described in this example were monitoredvia LCMS and HPLC as described in this section. Reaction products werecharacterized using the same conditions.

[0197] LCMS was performed on a Finnigan LCQ-Classic mass spectrometerwith a Waters Alliance HPLC used as the inlet device or on a ThermoquestLCQ-Duo mass spectrometer with a Thermoquest Surveyor HPLC as the inletdevice. The columns used were either a Waters Symmetry C18 (4.6×50 mm)or a Vydac C4 (4.6×50 mm). A flow rate of 0.75 mL/min was used with alinear gradient between Buffer A (0.5% acetic acid in water) and BufferB (0.5% acetic acid in acetonitrile).

[0198] HPLC was performed on an HP 1090 photo diode array HPLC or on anAgilent 1100 photo diode array HPLC. The columns used were the same asfor the LCMS analysis. A flow rate of 1.0 mL/min was used with a lineargradient between Buffer A (60 mM Et₃N-HOAc pH 6.0) and Buffer B (90%acetonitrile and 10% Buffer A).

[0199] 5.2 Preparation of Amide AmB Derivatives of Formula (II)

[0200] 5.2.1 Preparation of Sugar Amine Free Bases

[0201] Glucosamine, galactosamine and mannosamine were purchased ashydrochloride salts. For use in the preparation of amides as describedin Section 5.2.2 (infra), the free base form of the amines are required.The free base amines were prepared using ion exchange chromatography asdescribed below, illustrated with glucosamine hydrochloride as aspecific example. The other sugar amine free bases were prepared byidentical methods.

[0202] Anion exchange resin (Bio-Rad AG 3-X4, 100-200 mesh) was loadedinto a column (25 g) and washed with methanol. Sodium hydroxide solution(1 M, 50 mL) was added and the resin was washed with water until theeluant attained pH 7. A solution of glucosamine hydrochloride (5.0 g)dissolved in 50 mL water was added to the resin followed by elution with300 mL water. The water was lyophilized to give the amino sugar freebase.

[0203] 5.2.2 Preparation of Amides

[0204] Compounds 123, 124, 125 and 126 were synthesized as illustratedin Scheme (II), supra. Briefly, AmB (100 mg, 0.11 mmol) was dissolved in19.65 mL dimethylformamide in a round bottom flask. Triethylamine (100mL, 1.1 mmol) was added and the solution was stirred at room temperaturefor 10 min. The appropriate amine in free base form (see Section 5.2.1,supra; see also TABLE 2; 1.1 mmol) was then added, thoroughly mixed, andsonicated for 1 min, followed by the addition of diphenylphosphorylazide(250 mL, 1.1 mmol). The reaction was allowed to stand at roomtemperature for 6 hrs. The formation of the glycosylamide AmB productwas determined by analytical HPLC (Waters Symmetry C-18, 3.5 mm, 4.6×50mm) using a linear gradient of 30% to 45% of solvent B over 10 min at aflow rate of 1.1 mL/min, UV=365 nm, and 410 nm (solvent A: 100 mMtriethylammonium acetate pH 7/water; solvent B: 90% Acetonitrile/100 mMtriethylammonium acetate pH 7). The reaction mixture was then diluted bya factor of ten with 100 mM triethylammonium acetate pH 6 and thedesired product was purified by preparative reverse-phase HPLC usinglinear gradient of 25% to 45% of solvent B over 60 min at a flow rate of6 mL/min, UV=340 nm (solvent A: 100 mM triethylammonium acetate pH6/water; solvent B: 90% acetonitrile/100 mM triethylammonium acetate pH6). Fractions containing the amide product were pooled and the resultantaqueous solution was lyophilized several times to dry the product andremove excess triethylammonium salts. The structures were confirmed bymass spectra, and the purity was confirmed by HPLC analysis.

[0205] The major mass spec peak (MH⁺) for each compound synthesized isas follows: Compound 123, MH+=1086; Compound 124, MH+=1086; Compound125, MH+=1086; Compound 126, MH+=1087.

[0206] 5.3 Two-Pot Synthesis of AmB Amide Derivatives of Formula (I)

[0207] Certain amide derivatives according to formula (I) weresynthesized according to the “two-pot” method illustrated in Scheme (I),supra. Briefly, AmB was first reacted with a reducing sugar underAmadori rearrangement conditions using the procedure described inFalkowski et al., 1975, “N-Glycosyl Derivatives of Polyene MacrolideAntibiotics,” J. Antibiot. 28:244. The resultant glycosylatedintermediate was then amidated using standard methods. The specificprocedures used for each step of the two-pot synthesis are providedbelow (the Amadori rearragngement is illustrated with reference toα-D-Glucose). Compounds 100-122,127-130,132-136 and 140-144 weresynthesized according to this method.

[0208] 5.3.1 Amadori Rearrangement

[0209] AmB (800 mg, 0.86 mmol) was dissolved in 16 mL anhydrousdimethylformamide (DMF) and stirred for 5 min. α-D-Glucose (316 mg, 1.72mmol) was added. The resulting solution was sealed to the atmosphere andplaced in a water bath equilibrated to 37° C. for 6 hr. The reaction wasfollowed by LCMS until judged complete. The reaction product was useddirectly in the amidation reaction, described infra, withoutpurification. The corresponding D-galactose, D-maltose, D-cellobiose andα-D-galactose glycosylated rearrangement products were obtained usingthe same procedure, but substituting D-galactose (315 mg, 1.72 mmol),D-maltose hydrate (620 mg, 1.72 mmol), D-cellobiose (588 mg, 1.72 mmol)and α-D-lactose hydrate (620 mg, 1.72 mmol), respectively, forα-D-glucose.

[0210] 5.3.2 Amidation

[0211] To make the corresponding amides, diphenylphosphorylazide (110μL, 0.92 mmol), triethylamine (128 μL; 0.92 mmol) and the amines fromTABLE 2, infra (0.92 mmol), were added to the glycosylated Amadorirearrangement products (crude DMF solution) from Section 5.3.1, supra (2mL, ca. 100 mg, 0.1 mmol). The solutions were stirred at roomtemperature and monitored by LCMS. After 4 hr, the products wereprecipitated with 50 mL of cold diethyl ether. After centrifugation andremoval of the ether, the products were dissolved in 40 mL water andlyophilized to give yellow powders. The compound numbers (Cmpds.), aminereactants and reducing carbohydrates used to synthesize the compoundsare noted in TABLE 2, infra. The structures of the compounds synthesizedare also provided in the Detailed Description of the Invention, supra.

[0212] 5.4 One-Pot Synthesis of AmB Amide Derivatives of Formula (I)

[0213] Certain amide derivatives according to formula (I) weresynthesized according to the “one-pot” method illustrated in Scheme(III), supra, using either PyBroP or HBTU as the coupling reagent. Eachmethod is illustrated below with Compound 114. The other compoundssynthesized according to each method follow the reaction details.

[0214] 5.4.1 PyBroP One-Pot Synthesis

[0215] To a suspension of AmB (500 mg; 0.514 mmol) in 8 mL of DMF wasadded D-galactose (107 mg; 0.595 mmol). The reaction mixture was stirredfor 72 hr at 50° C. The mixture was cooled to 0° C., DIEA (0.377 mL;2.164 mmol) and PyBroP (504 mg; 1.082 mmol) were added and the resultantmixture was stirred for 10 min. 1-(2-Amino ethyl)piperidine (0.231 mL;1.623 mmol) was added, the mixture warmed to 25° C. and stirred for anadditional 1.5 hr. The mixture was then suspended in 50 mL ether,filtered through a fritted funned and the collected precipitate waswashed with ether (50 mL×2) and acetonitrile (50 mL×3) and dried underhouse vacuum for 1 hr to yield 640 mg of Compound 114.

[0216] Compounds 103, 104, 107, 110, 114, 133 and 135 were synthesizedby the same procedure, substituting the appropriate amounts of reducingcarbohydrate and amine.

[0217] 5.4.2 HBTU One-Pot Synthesis

[0218] To a suspension of AmB (500 mg; 0.514 mmol) in 8 mL of DMF wasadded D-galactose (107 mg; 0.595 mmol). The reaction mixture was stirredfor 72 hr at 50° C. The mixture was cooled to 0° C., DIEA (0.377 mL;2.164 mmol) and HBTU (410 mg; 1.082 mmol) were added and the resultantmixture was stirred for 10 min. 1-(2-Amino ethyl)piperidine (0.231 mL;1.623 mmol) was added, the mixture warmed to 25° C. and stirred for anadditional 1.5 hr. The mixture was then suspended in 50 mL ether,filtered through a fritted funned and the collected precipitate waswashed with ether (50 mL×2) and acetonitrile (50 mL×3) and dried underhouse vacuum for 1 hr to yield 660 mg of Compound 114.

[0219] 5.5 Alternative Amadori Rearrangement Conditions

[0220] The compounds of the invention may also be made using thealternative Amadori rearrangement conditions described below. This routeutilizes DMPU solvent and a total of 1.1 equiv reducing carbohydrateadded to the reaction mixture in three equal aliguots. It can be used inconnection with the two-pot synthesis or the one-pot synthesis.

[0221] To a solution of AmB (1 equiv) dissolved in 400 mL ofN,N′-dimethylpropylene urea (“DMPU”) at a temperature in the range of30° C. to 55° C., typically about 45° C., is added reducing carbohydrate(0.367 equiv). The mixture is stirred for about 1-3 hr, typically about1.35 hr, and another aliquot of reducing carbohydrate (0.367 equiv) isadded. The mixture is again stirred for 1.35 hr and a final aliquot ofreducing carbohydrate (0.367 equiv) added. This resulting mixture isthen stirred for an additional 18 hr at 45° C., and the resultantAmadori rearrangement product isolated as previously described.

[0222] 5.6 Preparation of Aspartate Salts

[0223] A polyene macrolide derivative is mixed with one equivalent ofaspartic acid and then water is added to a concentration of 50 mg/mL. Ifa clear solution does not result then small amounts of dimethylsulfoxideare added to provide a clear yellow solution, which is frozen andlyophilized to obtain a yellow powder. The yellow powder is dissolved inwater, frozen and lyophilized again to obtain the aspartate salt.

[0224] 5.7 Formulation of Polyene Macrolide Derivatives

[0225] The polyene macrolide amide derivatives of the invention, and inparticular the aspartic acid salts thereof, possess good watersolubility. Most of the compounds of the invention, particularly theaspartate salts, can be dissolved in 5% aq. mannitol (e.g., OSMITROL,Baxter Healthcare, Deerfield, Ill.) to produce yellow, isotonicsolutions. Compounds that do not readily dissolve in OSMITROL may bedissolved first in a 0.1% aq. lactic acid solution at pH of about 2.0.For intraperitoneal injections, propylene glycol can be added in smallquantities, up to about 10% (v/v), if the polyene macrolide derivativedoes not dissolve in the lactic acid. A 4.9% aq. mannitol solution(buffered with acetic acid to pH of 5.0) is then added to form a yellow,isotonic solution. TABLE 2 Sugar Used in Amadori Rearrangement AmineUsed in None α-D-Glucose D-Galactose D-Maltose D-Cellobiose α-D-LactoseAmidation Cmpd. Cmpd. Cmpd. Cmpd. Cmpd. Cmpd. A

100 108 138 B

101 109 116 C

102 132 139 D

126 103 110 117 141 E

104 111 118 144 F

105 112 119 G

106 113 140 143 146 H

107 114 120 142 I

115 121 J

122 K

145 L

127 133 M

128 134 N

129 135 147 O

130 136 P

131 137 Q

123 R

124 S

125

6. EXAMPLE In Vitro Data

[0226] For each compound synthesized, Minimum Inhibitory Concentrations(MICs) against C. albicans were determined as described in ReferenceMethod for Broth Dilution Antifungal Susceptibility Testing of Yeasts,Approved Standard, NCCLS document M27-A (ISBN 1-56238-328-0), NCCLS, 940West Valley Road, Suite 1400, Wayne, Pa. 19087, 1997. The MICs (inμg/mL) of the various compounds are provided in TABLE 3, infra). Allcompounds tested exhibited MICs of 8 μg/mL or less, and many exhibitedNEC comparable to AmB (0.125-0.25 μg/mL) and AME (0.75 μg/mL onaverage).

7. EXAMPLE In Vivo Data

[0227] A variety of the compounds synthesized as described in Section 5,supra, were tested in various in vivo assays, as described below. Theresults of the various assays are provided in TABLE 3, presented at theend of this section. In all of the in vivo assays described, the testcompounds, in the form of the aspartate salts, were formulated in anaqueous vehicle (5% mannitol in sterile water; OSMITROL, BaxterHealthcare, Deerfield, Ill.), followed by appropriate dilutions. Theformulations were prepared fresh before dosing.

[0228] AmB and AME were also tested for comparison. For AmB, thecommercially available FUNGIZONE (Bristol-Meyers Squibb Co.) was used.Reported values are based upon the weight of the AmB active ingredient,not on the basis of the weight of the FUNGIZONE powder. AME wasformulated in OSMITROL as decribed for the test compounds.

[0229] 7.1 Determination of Maximum Non-Lethal Dose

[0230] Maximum non-lethal dose (NLD) data were obtained for selectedcompounds. A single intravenous dose was administered to each of fourmice (Swiss-Webster male mice; Simonsen Laboratories, Gilroy, Calif.) inseveral groups. The doses selected were based upon the NLDs of compoundsof similar structures. After review of findings, higher or lower doseswere administered and a NLD was determined. The results are tabulated inTABLE 3, below (NLD Acute Mice). The values for AmB and AME are providedfor comparison.

[0231] Each compound evaluated in mice by the intravenous route ofadministration had a lower order of acute toxicity than AmB.

[0232] 7.2 Determination of ED₅₀: 5 Day Kidney Bioburden Evaluation inImmunocompetent Mice Challenged with C. Albicans

[0233] Various polyene macrolide amide derivatives of the invention wereevaluated for their protective effect when administeredintraperitoneally once daily for 5 days to immunocompetent micechallenged with live C. albicans.

[0234]C. albicans was grown on Sabouraud dextrose agar (SDA) plates for48 hr. Colonies were harvested and washed twice by centrifugation inphosphate-buffered saline (PBS). The washed pellets were resuspended inPBS and diluted to achieve the desired number of fungi per milliliter. Aportion of the inoculum was plated on SDA plates to determine thecolony-forming units (CFU) per milliliter.

[0235] Swiss-Webster mice (Simonsen Laboratories, Gilroy, Calif.) wereadministered by intravenous injection a single inoculum (0.2 mL)containing an estimated 2.0×10⁵ CFU of C. albicans. The day ofinoculation was designated as study day 0. Beginning three days afterinjection, mice were treated intraperitoneally with vehicle alone orformulated test compound once daily for 5 consecutive days.

[0236] The mice were observed daily from study day 0 through study day10 (3 days after the last dose). Mice found dead or euthanized inextremis were removed from the study without further processing.Surviving mice were euthanized by inhalation of CO₂ on study day 10 andboth kidneys from each mouse were removed. Each pair of removed kidneyswas homogenized in 3 mL of ice-cold PBS containing 10% glycerol. Aportion of each homogenate was used to determine log₁₀ CFU per kidneyusing SDA plates (48 hours of incubation at 30° C.). Kidney log₁₀ CFUdata were analyzed using a t-test. The criterion for a statisticallysignificant response was set at p<0.01.

[0237] The results are tabulated in TABLE 3 (ED₅₀ mice 5 day). Forcomparison, the results for AmB and AME are also provided. Efficacy wasdefined for individual animals as a greater than 2 log₁₀ reduction inkidney CFU relative to the mean vehicle control value. The ED₅₀ for thetest compound was determined as the dosage at which 50% of the animalsexhibited a 2 log₁₀ reduction in kidney CFU relative to the mean vehiclecontrol value.

[0238] When administered intraperitoneally at a dose of 2, 4, 8 or 16mg/kg/day, AmB was effective in 20%, 60%, 100% and 78% of mice,respectively. The ED₅₀ for mice treated with AmB intraperitoneally inthis model was calculated to be 3 mg/kg/day. Many of the compoundstested exhibited ED₅₀s comparable to AmB in this assay.

[0239] 7.3 Determination of ED₅₀: 7 Day Kidney Bioburden Evaluation onNeutropenic Mice Challenged with C. Albicans

[0240] Various polyene macrolide amide derivatives of the invention wereevaluated for their protective effect when administeredintraperitoneally once daily for 7 days to neutropenic mice challengedwith live C. albicans.

[0241] Swiss-Webster mice (Simonsen Laboratories, Gilroy, Calif.) weremade neutropenic by intravenous administration of 5-fluorouracil (150mg/kg) on study day −1. On study day 0, all mice received by intravenousinjection a single inoculum (0.2 mL) containing as estimated 5.0×10⁴ CFUof C. albicans. The day of inoculation was designated as study day 0.Beginning the day after infection (on study day 1), mice were treatedintraperitoneally with vehicle alone or formulated test compound, oncedaily for 7 consecutive days.

[0242] The mice were observed daily from study day 1 through study day 8(1 day after the last dose). Mice found dead or euthanized in extremiswere removed from the study without further processing. Surviving micewere euthanized by inhalation of CO₂ on study day 8 and both kidneysfrom each mouse were removed. Each pair of removed kidneys washomogenized in 3 mL of ice-cold PBS containing 10% glycerol. A portionof each homogenate was used to determine log₁₀ CFU per kidney using SDAplates (48 hours of incubation at 30° C.). Kidney log₁₀ CFU data wereanalyzed using a t-test. The criterion for a statistically significantresponse was set at p<0.01.

[0243] The results are tabulated in TABLE 3 (ED₅₀ mice 7 day). Forcomparison, the results for AmB and AME are also provided. Efficacy wasdefined for individual animals as a greater than 2 log₁₀ reduction inkidney CFU relative to the mean vehicle control value. The ED₅₀ for thetest compound was determined as the dosage at which 50% of the animalsexhibited a 2 log₁₀ reduction in kidney CFU relative to the mean vehiclecontrol value.

[0244] 7.4 Two Week Range-Finding Toxicity Study in Mice

[0245] Various polyene macrolide amide derivatives of the invention wereevaluated to characterize their potential toxicity when administeredsubchronically by intravenous injection.

[0246] CD-1® mice (Charles River Laboratories, Portage, Mich.) wereacclimated to laboratory conditions for at least 5 days before the startof dosing. During the acclimation period, the general condition of themice was evaluated, and those considered healthy were used. The micewere randomly assigned to treatment groups using a stratified bodyweight regimen. Each treatment group consisted of 3 male and 3 femalemice. The mice were housed in a room with a controlled environment andprovided rodent chow and water ad libitum.

[0247] On the first day of dosing, the mice were approximately 7 to 9weeks old and weighed 22 to 30 g. Doses were administered by intravenousinjection once daily on study days 1 to 5 and 8 to 12. Dosages wereselected based upon data from efficacy and toxicology studies, andranged from 1.25 to 30 mg/kg/day. Clinical observations were recorded0.5 hr after dosing on days 1, 3, 5, 8, 10, 12 and 15, and on any day achange was observed. Body weights were recorded prior to dosing and onstudy days 8 and 12, and on study day 15 at the completion of thein-life phase of the study.

[0248] Animals that were found in a moribund/deteriorating clinicalstate were euthanized (animal designated killed in extremis) byinhalation of CO₂. The day of death for any animal found dead or killedin extremis was recorded. Any animal that died prematurely was removedfrom the study without further processing.

[0249] One day after completion of dosing (study day 16), all survivingmice were euthanized by inhalation of CO₂. Their kidneys were preservedfor histopathologic examination. The maximum no-observed-adverse-effectlevel (NOAEL) was determined on the basis of clinical andhistopathologic findings. Results are tabulated in TABLE 3. The valuesfor AmB and AME are provided for comparison.

[0250] 7.5 Ascending Single-Dose Nephrotoxicity Study in Rabbits

[0251] Various polyene macrolide amide derivatives of the invention wereevaluated to determine the intravenous dose of administered compoundthat produces nephrotoxicity in rabbits. Nephrotoxicity was monitored bymeasuring urine volume and blood urine nitrogen (BUN) and creatinine.

[0252] Female New Zealand white rabbits (Hra: (NZW) SPF; Covance,Richmond, Calif.) were acclimated to laboratory conditions for 9 daysbefore the start of dosing. During the acclimation period, the rabbitswere acclimated to restrainers on four separate days. The duration ofrestraint on the first day of acclimation did not exceed 5 min. Theduration of restraint on the last day of acclimation was 50 min. Duringthe study, the animals were restrained for no more than 60 min per day.

[0253] The rabbits were housed in a room with a controlled environmentand placed on a restricted diet of 125 g rabbit chow per day. Water wasprovided ad libitum.

[0254] Ascending single doses were administered by intravenous infusioninto an ear vein at a rate of 0.6 mL/min. Each ascending dose wasfollowed by a non-dosing observation period of at least two days. Theclinical condition of each rabbit was observed daily for generalcondition and availability of food and water. Body weights were recordedbefore each dose, as well as 2 days after the last dose. Blood sampleswere collected from the rabbits two days prior to the first dose (studyday −2) and approximately 24 hr after each dose and analyzed for BUN andcreatinine. Urine was collected for two 24-hour periods prior to theinitiation of dosing and for the 24-hour period following eachtreatment. Necropsy was not performed.

[0255] Body weight, urine output, BUN and creatinine data were evaluatedstatistically using a one-tailed Mann-Whitney test. The criterion forstatistical significance was set at p≦0.05. The maximumno-observed-adverse-effect level (NOAEL) of nephrotoxicity wasdetermined on the basis of the above measurements. Results are tabulatedin TABLE 3. The values for AmB and AME are provided for comparison.

[0256] 7.6 Determination of ED₅₀ in a Mouse Survival Model

[0257] Swiss-Webster mice (Simonsen Laboratories, Gilroy, Calif.) aremade neutropenic by intravenous administration of 5-fluorouracil (150mg/kg) on study day −1.

[0258] On study day 0, all mice receive by intravenous injection asingle inoculum (0.5 mL) containing as estimated 5.0×10⁴ CFU of C.albicans. The day of inoculation is designated as study day 0. Atapproximately 4 hr post-injection, mice are treated intraperitoneallywith test compound once daily for 7 consecutive days.

[0259] The mice are observed daily from study day 0 through study day28. Mice found dead or euthanized due to moribund condition are removedfrom the study. Surviving mice are euthanized by inhalation of CO₂ onstudy day 28 and removed from the study. Moribund mice are recorded asdead the day after euthanasia.

[0260] The ED₅₀ for the test compound was determined as the dosage atwhich 50% of the animals survived.

[0261] 7.7 Kidney Bioburden Evaluation in Immunosupressed MiceChallenged with Candida Glabrata

[0262]C. glabrata is grown in 20 mL SBA broth (250 mL Ehrlenmeyer) for24 hr at 37° C. in a stationary incubator. The suspension is washedtwice by centrifugation in PBS. The washed pellets are resuspended inPBS and diluted to achieve the desired number of fungi per milliliter. Aportion of the inoculum is plated on SDA plates to determine thecolony-forming units (CFU) per milliliter.

[0263] Black female C57 mice are immunosupressed by administrationintraperitoneally of cyclophosphamide (100 mg/kg) on study days −3, 0,3, 6 and 9. On study day 0, all mice receive by intravenous injection asingle inoculum (0.2 mL) containing an estimated 1.5×10⁸ CFU of C.glabrata. Beginning 4 days after inoculation, mice are treatedintraperitoneally with either vehicle control or test compoundformulation once daily for 5 consecutive days.

[0264] The mice are observed daily through study day 11 (3 days afterthe last dose). Mice found dead or euthanized in extremis are removedfrom the study without further processing. Surviving mice are euthanizedby inhalation of CO₂ on study day 11 and both kidneys from each mouseare removed. Each pair of removed kidneys is homogenized in 3 mL ofice-cold PBS containing 10% glycerol. A portion of each homogenate wasused to determine log₁₀ CFU per kidney using SDA plates (48 hours ofincubation at 37° C.). Kidney log₁₀ CFU data are analyzed using at-test. The criterion for a statistically significant response was setat p<0.01.

[0265] Efficacy is defined for individual animals as a greater than 2log₁₀ reduction in kidney CFU relative to the mean vehicle controlvalue. The ED₅₀ for test compounds is determined as the dosage at which50% of the animals exhibit a 2 log₁₀ reduction in kidney CFU relative tothe mean vehicle control value. TABLE 3 NLD ED₅₀ ED₅₀ NOAEL (mg/kg)NOAEL Compound (mg/kg) (mg/kg) (mg/kg) 2 week mice (mg/kg) No. MIC(μg/ML) acute mice mice 5 day mice 7 day histological clinical acuterabbit AMB 0.125-0.25 1.5 3 0.3 0.85 0.5 0.375 AME 0.75 60 13.18.3 >32 >32 7.5 123 5.3 32 124 1.0 16 125 1.0 8 100 2.0 101 4.0 102 2.7103 1.1 10 15 104 2.0 20 9 3.1 <10 10 7.5 105 2.0 106 2.0 <15 3 6 3 1072.0 >40 28 110 1.0 10 15 108 4.0 113 3.3 7.5 2 1.25 >5 109 2.0 111 2.720 7 7 <7 132 112 114 4.0 40 9 6.1 9 9 7.5 115 116 8.0 139 117 8.0 1185.3 119 8.0 120 8.0 121 8.0 122 8.0 140 141 142 143 144 138 127 128 129130 133 15 4.1 NA <5 134 1.0 30 >30 135 6.3 10 1.6 >5 1.25 136 0.4 >1540.2 146 2.0 <7.5 147 2.0 <7.5 131 4.0 5 137 2.0 5 145 1.0 20 3 123 5.332 124 1.0 16 125 1.0 8 126 3.0

[0266] Having been described, the present invention is not to be limitedin scope by the exemplified embodiments, which are intended asillustrations of single aspects of the invention, and any compositionsand methods which are functionally equivalent are within the scope ofthe invention. Indeed, various modifications of the invention inaddition to those described above will become apparent to those skilledin the art from the foregoing description and accompanying drawings.Such modifications are intended to fall within the scope of the appendedclaims.

We claim:
 1. A compound according to structural formula (I):

including the pharmaceutically acceptable salts thereof, wherein:N—R¹—C(O) is a polyene macrolide backbone; CH₂—R² is a carbohydrateresidue, where the illustrated CH₂ is derived from the anomeric carbonof a terminal carbohydrate saccharide and R² represents the remainder ofthe carbohydrate; either: (i) R⁶ is selected from the group consistingof hydrogen, non-polar substituent and water-solubility increasingsubstituent and R⁷ is a water-solubility increasing substituent; or (ii)R⁶ and R⁷, taken together with the amide nitrogen to which they arebonded, form a saturated or unsaturated ring which optionally includesone or more of the same or different ring heteroatoms and which isoptionally substituted at one or more ring carbon or heteroatoms withthe same or different polar or non-polar substituents or combinationsthereof; and R¹⁴ is hydrogen or alkyl.
 2. The compound of claim 1 inwhich polyene macrolide backbone N—R¹—C(O) is derived from amphotericinB or nystatin.
 3. The compound of claim 1 in which R¹⁴ is hydrogen. 4.The compound of claim 1 in which: either: (i) R⁶ is selected from thegroup consisting of hydrogen, (C₁-C₆) alkyl, (C₁-C₆) alkyl substitutedwith one or more of the same or different R¹⁰ groups,—[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—NR¹⁵R¹⁶—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—NR¹⁵R¹⁶,[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—R¹⁷ and—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—R¹⁷ and R⁷ is selected from the groupconsisting of (C₁-C₆) alkyl substituted with one or more of the same ordifferent R¹⁰ groups, —[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—NR¹⁵R¹⁶,—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—NR¹⁵R¹⁶,—[(CH₂)_(n)—NH]_(p)—(CH₂)_(n)—R¹⁷ and—NH—[(CH₂)_(n)—NH]_(p)—(CH₂)_(p)—R¹⁷; or (ii) R⁶ and R⁷, taken togetherwith the amide nitrogen atom to which they are bonded, form a 5- or6-membered saturated or unsaturated ring which optionally includes oneor more of the same or different additional heteroatoms selected fromthe group consisting of O, N, NH and S and which is optionallysubstituted at one or more ring carbon or heteroatoms with the same ordifferent substituents selected from the group consisting of R¹⁰,(C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰, (C₅-C₆) aryl, phenyl, 6-to 9-membered arylalkyl and benzyl; each R¹⁰ is independently selectedfrom the group consisting of —OH, ═O (oxo), —NH₂ (amino), ═NH (imino),—C(═NH)—NH₂ (amidino) and —NH—C(═NH)—NH₂ (guanidino); either: (i) R¹⁵and R¹⁶ are each independently selected from the group consisting ofhydrogen, (C₁-C₆) alkyl and (C₁-C₆) alkyl independently substituted withone or more of the same or different R¹⁰ groups; or (ii) R¹⁵ and R¹⁶,taken together with the nitrogen atom to which they are bonded, form a5- or 6-membered saturated or unsaturated ring which optionally includesone or more of the same or different additional heteroatoms selectedfrom the group consisting of O, N, NH and S and which is optionallysubstituted at one or more ring carbon or heteroatoms with the same ordifferent substituents selected from the group consisting of R¹⁰,(C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰, (C₅-C₆) aryl, phenyl, 6-to 9-membered arylalkyl and benzyl; R¹⁷ is a 5- or 6-membered saturatedor unsaturated ring including one or more of the same or differentheteroatoms selected from the group consisting of O, N, NH and S andwhich is optionally substituted at one or more ring carbon orheteroatoms with the same or different substituents selected from thegroup consisting of R¹⁰, (C₁-C₆) alkyl, (C₁-C₆) alkoxy, —(CH₂)_(r)—R¹⁰,(C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl and benzyl; each n isindependently an integer from 1 to 6; and each p is independently aninteger from 0 to
 6. 5. The compound of claim 4 in which R⁶ and R⁷ aredefined according to alternative (i).
 6. The compound of claim 4 inwhich R⁶ and R⁷ are defined according to alternative (ii).
 7. Thecompound of claim 1 or 4 which has one or more features selected fromthe group consisting of: N—R¹—C(O) is a polyene backbone derived fromAmB or nystatin; CH₂—R² is a mono-, di- or oligosaccharide; R⁶ ishydrogen; and R¹⁴ is hydrogen.
 8. The compound of claim 4 in which: R⁶is hydrogen; R⁷ is selected from the group consisting Of —NH—NR¹⁵R¹⁶,—(CH₂)_(n)—NR¹⁵R¹⁶, —(CH₂)_(n)—R¹⁷ and (C₁-C₆) alkyl substituted withone or more amino or hydroxyl groups; R¹⁵ and R¹⁶, taken together withthe nitrogen atom to which they are bonded, form a 5- or 6-memberedsaturated or unsaturated ring which optionally includes one or moreadditional heteroatoms selected from the group consisting of O, S, N andNH and/or which is optionally substituted at one or more ring carbon orheteroatoms with the same or different R¹⁰, (C₁-C₆) alkyl, (C₁-C₆)alkoxy, (C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl or benzylgroups; R¹⁷ is a 5- or 6-membered heteroaryl which is optionallysubstituted with one or more of the same or different (C₁-C₆) alkyl,(C₁-C₆) alkoxy, (C₅-C₆) aryl, phenyl, 6- to 9-membered arylalkyl orbenzyl groups.
 9. The compound of claim 4 in which: R⁶ and R⁷, takentogether with the nitrogen atom to which they are bonded, form a 5- to6-membered cycloheteroalkyl ring which is optionally substituted withone or more substituent selected from the group consisting of (C₁-C₆)alkyl, (C₁-C₆) alkoxy, —(CH₂)_(n)—R¹⁰, (C₅₋₆) aryl, phenyl, 6- to9-membered arylalkyl and benzyl; and R¹⁰ is amino or hydroxy.
 10. Thecompound of claim 1 in which —CH₂—R² is a mono-, di- or oligosaccharide.11. The compound of claim 10 in which —CH₂—R² is an Amadorirearrangement product of a reducing carbohydrate selected from the groupconsisting of glucose, galactose, maltose, cellobiose and lactose. 12.The compound of claim 1 or 11 in which substituent NR⁶R⁷ is contributedby any of the amines listed in TABLE
 2. 13. The compound of claim 1which is selected from the group consisting of Compounds 100, 101, 102,103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116,117, 118, 119, 120, 121, 122, 127, 128, 129, 130, 131, 132, 133, 134,135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146 and
 147. 14.A compound according to structural formula (II):

including the pharmaceutically acceptable salts thereof, wherein:N—R¹—C(O) and R¹⁴ are as previously defined in claim 1; R³ is hydrogen,a non-polar substituent or a water-solubility increasing substituent; R⁴is hydrogen or alkyl; and R⁵ is a water-solubility increasingsubstituent selected from the group consisting of polyhydroxylatedalkyl, monosaccharide, disaccharide and oligosaccharide.
 15. Thecompound of claim 14 in which polyene macrolide backbone N—R¹—C(O) isderived from amphotericin B or nystatin.
 16. The compound of claim 14 inwhich R⁴ is hydrogen.
 17. The compound of claim 14 in which R³ is awater-solublity increasing substituent selected from the groupconsisting of polyhydroxylated alkyl, monosaccharide, disaccharide andoligosaccharide
 18. The compound of claim 14 in which R³ is hydrogen orlower alkyl.
 19. The compound of claim 14 in which R⁵ is selected fromthe group consisting of glucosyl, galactosyl and mannosyl.
 20. Thecompound of claim 14 which is selected from the group consisting ofCompounds 123, 124, 125 and
 126. 21. A pharmaceutical compositioncomprising a compound according to claim 1 or claim 14 and apharmaceutically-acceptable carrier, excipient or diluent.
 22. A methodof inhibiting the growth of a fungus comprising contacting the funguswith an amount of a compound according to claim 1 or claim 14 effectiveto inhibit the growth of the fungus.
 23. A method of treating orpreventing a fungal infection in a subject comprising administering to asubject an amount of a compound according to claim 1 or claim 14effective to treat or prevent the fungal infection.
 24. The method ofclaim 23 in which the subject is a human, an animal or a plant.
 25. Themethod of claim 23 in which the infection is a topical infection. 26.The method of claim 23 in which the infection is a systemic infection.27. A method of making a polyene macrolide amide derivative, comprisingthe steps of: reacting a parent polyene macrolide with a reducingcarbohydrate under Amadori rearrangement conditions to yield an Amadorirearrangement product; and amidating the Amadori rearrangement productwith an amine reagent of the formula HNR⁶R⁷, where R⁶ and R⁷ are asdefined in claim 1, to yield the polyene amide macrolide derivative. 28.The method of claim 26 which further includes the step of N-alkylatingthe parent polyene macrolide, the Amadori rearrangement product or theresultant polyene macrolide amide derivative.
 29. A method of making apolyene macrolide amide derivative, comprising the steps of: amidating aparent polyene macrolide with an amine reagent of the formula HNR⁶R⁷,where R⁶ and R⁷ are as defined in claim 1, to yield an amidated polyenemacrolide; and reacting the amidated polyene macrolide with a reducingcarbohydrate under Amadori rearrangement conditions to yield the polyenemacrolide amide derivative.
 30. The method of claim 28 which furtherincludes the step of N-alkylating the parent polyene macrolide, theamidated polyene macrolide or the resultant polyene macrolide amidederivative.
 31. The method of claim 26 or 28 in which the amidation stepis effected with an uronium salt or phosphonium salt coupling reagent.32. The method of claim 26 which is carried out in a single pot.
 33. Amethod of making a polyene macrolide amide derivative according tostructural formula (I):

wherein N—R¹—C(O), CH₂—R², R⁶, R⁷ and R¹⁴ are as defined in claim 1,comprising the steps of: reacting a polyene macrolide according tostructure (III):

with a reducing carbohydrate under Amadori rearrangement conditions toyield an Amadori rearrangement product; and amidating the Amadorirearrangement product with an amine reagent of the formula HNR⁶R⁷ toyield the polyene macrolide amide derivative of formula (I).
 34. Amethod of making a polyene macrolide amide derivative, comprising thesteps of: amidating a parent polyene macrolide with an amine reagent ofthe formula HNR³R⁵, where R³ and R⁵ are as defined in claim 14, to yieldthe polyene macrolide amide derivative.
 35. The method of claim 33 whichfurther includes the step of N-mono- or dialkylating the parent polyenemacrolide or the resultant polyene macrolide amide derivative.
 36. Themethod of claim 33 in which the amidation step is effective with anuronium salt or phosphonium salt coupling reagent.
 37. A method ofmaking a polyene macrolide amide derivative according to structuralformula (II):

wherein N—R¹—C(O), R³, R⁴, R⁵ and R¹⁴ are as defined in claim 14,comprising the steps of: reacting a polyene macrolide according tostructure (IV):

with an amine reagent of the formula HNR³R⁵, where R³ and R⁵ are asdefined in claim 14, to yield the polyene macrolide amide derivativeaccording to structural formula (II).
 38. The method of claim 36 inwhich the amidation step is effective with an uronium salt orphosphonium salt coupling reagent.
 39. The polyene macrolide amidederivative produce by the method of any one of claims 26, 28, 32, 33 or36.